pax_global_header00006660000000000000000000000064142327370040014514gustar00rootroot0000000000000052 comment=bdbba0a80cd13991295a73b53e9c6a862b87a311 kissplice-2.6.2/000077500000000000000000000000001423273700400135115ustar00rootroot00000000000000kissplice-2.6.2/AUTHORS000066400000000000000000000010031423273700400145530ustar00rootroot00000000000000 Vincent Lacroix (2011-) Leandro Lima (2014-) Gustavo Sacomoto (2011-2014) Pierre Peterlongo (2011-) Rayan Chikhi (2011-) Vincent Miele (2012-) Alice Julien-Laferrière (2012-2013) Camille Marchet (2013-) Audric Cologne François Gindraud (2022-) With the valuable help of Janice Kielbassa, Marie-France Sagot, David Parsons, Raluca Uricaru, Pavlos Antoniou and Gunter Roeth Contact: Vincent Lacroix , kissplice-users@lists.gforge.inria.fr kissplice-2.6.2/CMakeLists.txt000066400000000000000000000122471423273700400162570ustar00rootroot00000000000000# 3.1 required by: zlib targets, Threads (OpenMP target fallback) cmake_minimum_required(VERSION 3.1) project( kissplice VERSION 2.6.2 # Definition which is propagated through PROJECT_VERSION LANGUAGES CXX ) set(CMAKE_CXX_STANDARD 11) # Default to C++11 for C++ targets set(CMAKE_CXX_STANDARD_REQUIRED ON) # Fail if not supported (very old compiler at this point) # Build type, should be set by user. Useful predefined values (see `cmake -LH`): Debug, Release, RelWithDebInfo. # Default to non-persistent RelWithDebInfo (-g -O2) if no build type is specified. if(NOT CMAKE_BUILD_TYPE) # Not cached : this keeps the help string (`cmake -LH`) and user setting intact. set(CMAKE_BUILD_TYPE RelWithDebInfo) endif() # Cmake already adds -g in Debug and -O3 in Release, so no need to set them manually. # Enables common warnings for ALL targets. add_compile_options(-Wall) # Profiling with gprof requires -pg which is better to add manually with -DCMAKE_(C|CXX)_FLAGS=-pg # `perf` is a more modern replacement anyway and does not require special compilation. # Date for manpage and pdf doc string(TIMESTAMP CONFIGURE_DATE "%Y-%m-%d" UTC) ############################################################################### # structure and paths # kissplice is composed of: # - internal binaries which should go to /libexec/kissplice # - a main frontend "binary" which is a python script which should go to /bin # The frontend script expects this architecture to hold (with relative paths) to find the internal binaries. # For tests to work, this architecture is replicated in the temporary build directory wih = include(GNUInstallDirs) # GNU standard installation directories definitions (bin, libexec, and definitions for doc/man) # Path to internal binaries relative to prefix. This is used in install() for internal binaries, to let the cmake prefix handling work. set(RELATIVE_INTERNAL_BINDIR "${CMAKE_INSTALL_LIBEXECDIR}/kissplice") # Absolute path to build location of internal binaries. This is used to define RUNTIME_OUTPUT_DIRECTORY of internal binaries (build dir). set(BUILD_INTERNAL_BINDIR "${PROJECT_BINARY_DIR}/${RELATIVE_INTERNAL_BINDIR}") # Absolute path to build location of frontend binaries (kissplice script only for now) set(BUILD_BINDIR "${PROJECT_BINARY_DIR}/${CMAKE_INSTALL_BINDIR}") # Relative path from script to internal binaries directory, written to kissplice script file(RELATIVE_PATH KISSPLICE_BINDIR_TO_INTERNAL_BINDIR "${BUILD_BINDIR}" "${BUILD_INTERNAL_BINDIR}") # Complete the main script template to include the relative directory configure_file("${PROJECT_SOURCE_DIR}/kissplice.in.py" "${BUILD_BINDIR}/kissplice" @ONLY) install(PROGRAMS "${BUILD_BINDIR}/kissplice" DESTINATION "${CMAKE_INSTALL_BINDIR}") ############################################################################### # bcalm dependency # bcalm is a binary program needed for tests and after install. # It is packaged on debian, so default to using the system wide version (best practice for packaging kissplice). # The USE_BUNDLED_BCALM option can be used to locally build and ship an install of bcalm, placed in the internal binaries directory. # In either case a "bcalm" executable will be placed in internal binaries (bundled binary or a redirect script). # This simplifies kissplice.py which can always look in libexec/kissplice/bcalm. # This "bcalm" is placed in both build dir and install dir. option( USE_BUNDLED_BCALM "by default kissplice requires an already installed bcalm program ; enable this option to bundle a local bcalm with kissplice." OFF ) if(NOT USE_BUNDLED_BCALM) # Check system bcalm find_program(SYSTEM_BCALM_PATH NAMES bcalm) mark_as_advanced(SYSTEM_BCALM_PATH) if(NOT SYSTEM_BCALM_PATH) message( FATAL_ERROR "bcalm not found. install it with system packages,\ or provide path to a custom install with -DCMAKE_PROGRAM_PATH=,\ or use bundled bcalm version with -DUSE_BUNDLED_BCALM=TRUE" ) endif() message(STATUS "Found bcalm tool: ${SYSTEM_BCALM_PATH}") # Add a simple redirection to system bcalm in internal binaries, as kissplice expects bcalm there. configure_file("${CMAKE_CURRENT_SOURCE_DIR}/thirdparty/bcalm_redirect.in" "${BUILD_INTERNAL_BINDIR}/bcalm" @ONLY) else() message(STATUS "using bundled version of bcalm (will be compiled at make time)") include(ExternalProject) ExternalProject_Add( bundled_bcalm PREFIX bundled_bcalm GIT_REPOSITORY "https://github.com/GATB/bcalm.git" GIT_TAG v2.2.3 UPDATE_COMMAND "" # Prevent rebuilding bcalm everytime # Prevent install which is broken (vendored hdf5 ignores CMAKE_INSTALL_PREFIX). # Manually extract the bcalm binary to our internal binary build dir. This is run inside bcalm build dir. INSTALL_COMMAND "${CMAKE_COMMAND}" -E copy bcalm "${BUILD_INTERNAL_BINDIR}" ) endif() install(PROGRAMS "${BUILD_INTERNAL_BINDIR}/bcalm" DESTINATION "${RELATIVE_INTERNAL_BINDIR}") ############################################################################### # subdirs add_subdirectory(thirdparty) # kissreads add_subdirectory(modules) # various ks_* internal tools add_subdirectory(man) # manpage add_subdirectory(doc) # user guide enable_testing() add_subdirectory(tests) kissplice-2.6.2/COPYING000066400000000000000000001243761423273700400145610ustar00rootroot00000000000000CeCILL FREE SOFTWARE LICENSE AGREEMENT (english version, french version below) Notice This Agreement is a Free Software license agreement that is the result of discussions between its authors in order to ensure compliance with the two main principles guiding its drafting: * firstly, compliance with the principles governing the distribution of Free Software: access to source code, broad rights granted to users, * secondly, the election of a governing law, French law, with which it is conformant, both as regards the law of torts and intellectual property law, and the protection that it offers to both authors and holders of the economic rights over software. The authors of the CeCILL (for Ce[a] C[nrs] I[nria] L[ogiciel] L[ibre]) license are: Commissariat à l'Energie Atomique - CEA, a public scientific, technical and industrial research establishment, having its principal place of business at 25 rue Leblanc, immeuble Le Ponant D, 75015 Paris, France. Centre National de la Recherche Scientifique - CNRS, a public scientific and technological establishment, having its principal place of business at 3 rue Michel-Ange, 75794 Paris cedex 16, France. Institut National de Recherche en Informatique et en Automatique - INRIA, a public scientific and technological establishment, having its principal place of business at Domaine de Voluceau, Rocquencourt, BP 105, 78153 Le Chesnay cedex, France. Preamble The purpose of this Free Software license agreement is to grant users the right to modify and redistribute the software governed by this license within the framework of an open source distribution model. The exercising of these rights is conditional upon certain obligations for users so as to preserve this status for all subsequent redistributions. In consideration of access to the source code and the rights to copy, modify and redistribute granted by the license, users are provided only with a limited warranty and the software's author, the holder of the economic rights, and the successive licensors only have limited liability. In this respect, the risks associated with loading, using, modifying and/or developing or reproducing the software by the user are brought to the user's attention, given its Free Software status, which may make it complicated to use, with the result that its use is reserved for developers and experienced professionals having in-depth computer knowledge. Users are therefore encouraged to load and test the suitability of the software as regards their requirements in conditions enabling the security of their systems and/or data to be ensured and, more generally, to use and operate it in the same conditions of security. This Agreement may be freely reproduced and published, provided it is not altered, and that no provisions are either added or removed herefrom. This Agreement may apply to any or all software for which the holder of the economic rights decides to submit the use thereof to its provisions. Article 1 - DEFINITIONS For the purpose of this Agreement, when the following expressions commence with a capital letter, they shall have the following meaning: Agreement: means this license agreement, and its possible subsequent versions and annexes. Software: means the software in its Object Code and/or Source Code form and, where applicable, its documentation, "as is" when the Licensee accepts the Agreement. Initial Software: means the Software in its Source Code and possibly its Object Code form and, where applicable, its documentation, "as is" when it is first distributed under the terms and conditions of the Agreement. Modified Software: means the Software modified by at least one Contribution. Source Code: means all the Software's instructions and program lines to which access is required so as to modify the Software. Object Code: means the binary files originating from the compilation of the Source Code. Holder: means the holder(s) of the economic rights over the Initial Software. Licensee: means the Software user(s) having accepted the Agreement. Contributor: means a Licensee having made at least one Contribution. Licensor: means the Holder, or any other individual or legal entity, who distributes the Software under the Agreement. Contribution: means any or all modifications, corrections, translations, adaptations and/or new functions integrated into the Software by any or all Contributors, as well as any or all Internal Modules. Module: means a set of sources files including their documentation that enables supplementary functions or services in addition to those offered by the Software. External Module: means any or all Modules, not derived from the Software, so that this Module and the Software run in separate address spaces, with one calling the other when they are run. Internal Module: means any or all Module, connected to the Software so that they both execute in the same address space. GNU GPL: means the GNU General Public License version 2 or any subsequent version, as published by the Free Software Foundation Inc. Parties: mean both the Licensee and the Licensor. These expressions may be used both in singular and plural form. Article 2 - PURPOSE The purpose of the Agreement is the grant by the Licensor to the Licensee of a non-exclusive, transferable and worldwide license for the Software as set forth in Article 5 hereinafter for the whole term of the protection granted by the rights over said Software. Article 3 - ACCEPTANCE 3.1 The Licensee shall be deemed as having accepted the terms and conditions of this Agreement upon the occurrence of the first of the following events: * (i) loading the Software by any or all means, notably, by downloading from a remote server, or by loading from a physical medium; * (ii) the first time the Licensee exercises any of the rights granted hereunder. 3.2 One copy of the Agreement, containing a notice relating to the characteristics of the Software, to the limited warranty, and to the fact that its use is restricted to experienced users has been provided to the Licensee prior to its acceptance as set forth in Article 3.1 hereinabove, and the Licensee hereby acknowledges that it has read and understood it. Article 4 - EFFECTIVE DATE AND TERM 4.1 EFFECTIVE DATE The Agreement shall become effective on the date when it is accepted by the Licensee as set forth in Article 3.1. 4.2 TERM The Agreement shall remain in force for the entire legal term of protection of the economic rights over the Software. Article 5 - SCOPE OF RIGHTS GRANTED The Licensor hereby grants to the Licensee, who accepts, the following rights over the Software for any or all use, and for the term of the Agreement, on the basis of the terms and conditions set forth hereinafter. Besides, if the Licensor owns or comes to own one or more patents protecting all or part of the functions of the Software or of its components, the Licensor undertakes not to enforce the rights granted by these patents against successive Licensees using, exploiting or modifying the Software. If these patents are transferred, the Licensor undertakes to have the transferees subscribe to the obligations set forth in this paragraph. 5.1 RIGHT OF USE The Licensee is authorized to use the Software, without any limitation as to its fields of application, with it being hereinafter specified that this comprises: 1. permanent or temporary reproduction of all or part of the Software by any or all means and in any or all form. 2. loading, displaying, running, or storing the Software on any or all medium. 3. entitlement to observe, study or test its operation so as to determine the ideas and principles behind any or all constituent elements of said Software. This shall apply when the Licensee carries out any or all loading, displaying, running, transmission or storage operation as regards the Software, that it is entitled to carry out hereunder. 5.2 ENTITLEMENT TO MAKE CONTRIBUTIONS The right to make Contributions includes the right to translate, adapt, arrange, or make any or all modifications to the Software, and the right to reproduce the resulting software. The Licensee is authorized to make any or all Contributions to the Software provided that it includes an explicit notice that it is the author of said Contribution and indicates the date of the creation thereof. 5.3 RIGHT OF DISTRIBUTION In particular, the right of distribution includes the right to publish, transmit and communicate the Software to the general public on any or all medium, and by any or all means, and the right to market, either in consideration of a fee, or free of charge, one or more copies of the Software by any means. The Licensee is further authorized to distribute copies of the modified or unmodified Software to third parties according to the terms and conditions set forth hereinafter. 5.3.1 DISTRIBUTION OF SOFTWARE WITHOUT MODIFICATION The Licensee is authorized to distribute true copies of the Software in Source Code or Object Code form, provided that said distribution complies with all the provisions of the Agreement and is accompanied by: 1. a copy of the Agreement, 2. a notice relating to the limitation of both the Licensor's warranty and liability as set forth in Articles 8 and 9, and that, in the event that only the Object Code of the Software is redistributed, the Licensee allows future Licensees unhindered access to the full Source Code of the Software by indicating how to access it, it being understood that the additional cost of acquiring the Source Code shall not exceed the cost of transferring the data. 5.3.2 DISTRIBUTION OF MODIFIED SOFTWARE When the Licensee makes a Contribution to the Software, the terms and conditions for the distribution of the resulting Modified Software become subject to all the provisions of this Agreement. The Licensee is authorized to distribute the Modified Software, in source code or object code form, provided that said distribution complies with all the provisions of the Agreement and is accompanied by: 1. a copy of the Agreement, 2. a notice relating to the limitation of both the Licensor's warranty and liability as set forth in Articles 8 and 9, and that, in the event that only the object code of the Modified Software is redistributed, the Licensee allows future Licensees unhindered access to the full source code of the Modified Software by indicating how to access it, it being understood that the additional cost of acquiring the source code shall not exceed the cost of transferring the data. 5.3.3 DISTRIBUTION OF EXTERNAL MODULES When the Licensee has developed an External Module, the terms and conditions of this Agreement do not apply to said External Module, that may be distributed under a separate license agreement. 5.3.4 COMPATIBILITY WITH THE GNU GPL The Licensee can include a code that is subject to the provisions of one of the versions of the GNU GPL in the Modified or unmodified Software, and distribute that entire code under the terms of the same version of the GNU GPL. The Licensee can include the Modified or unmodified Software in a code that is subject to the provisions of one of the versions of the GNU GPL, and distribute that entire code under the terms of the same version of the GNU GPL. Article 6 - INTELLECTUAL PROPERTY 6.1 OVER THE INITIAL SOFTWARE The Holder owns the economic rights over the Initial Software. Any or all use of the Initial Software is subject to compliance with the terms and conditions under which the Holder has elected to distribute its work and no one shall be entitled to modify the terms and conditions for the distribution of said Initial Software. The Holder undertakes that the Initial Software will remain ruled at least by this Agreement, for the duration set forth in Article 4.2. 6.2 OVER THE CONTRIBUTIONS The Licensee who develops a Contribution is the owner of the intellectual property rights over this Contribution as defined by applicable law. 6.3 OVER THE EXTERNAL MODULES The Licensee who develops an External Module is the owner of the intellectual property rights over this External Module as defined by applicable law and is free to choose the type of agreement that shall govern its distribution. 6.4 JOINT PROVISIONS The Licensee expressly undertakes: 1. not to remove, or modify, in any manner, the intellectual property notices attached to the Software; 2. to reproduce said notices, in an identical manner, in the copies of the Software modified or not. The Licensee undertakes not to directly or indirectly infringe the intellectual property rights of the Holder and/or Contributors on the Software and to take, where applicable, vis-à-vis its staff, any and all measures required to ensure respect of said intellectual property rights of the Holder and/or Contributors. Article 7 - RELATED SERVICES 7.1 Under no circumstances shall the Agreement oblige the Licensor to provide technical assistance or maintenance services for the Software. However, the Licensor is entitled to offer this type of services. The terms and conditions of such technical assistance, and/or such maintenance, shall be set forth in a separate instrument. Only the Licensor offering said maintenance and/or technical assistance services shall incur liability therefor. 7.2 Similarly, any Licensor is entitled to offer to its licensees, under its sole responsibility, a warranty, that shall only be binding upon itself, for the redistribution of the Software and/or the Modified Software, under terms and conditions that it is free to decide. Said warranty, and the financial terms and conditions of its application, shall be subject of a separate instrument executed between the Licensor and the Licensee. Article 8 - LIABILITY 8.1 Subject to the provisions of Article 8.2, the Licensee shall be entitled to claim compensation for any direct loss it may have suffered from the Software as a result of a fault on the part of the relevant Licensor, subject to providing evidence thereof. 8.2 The Licensor's liability is limited to the commitments made under this Agreement and shall not be incurred as a result of in particular: (i) loss due the Licensee's total or partial failure to fulfill its obligations, (ii) direct or consequential loss that is suffered by the Licensee due to the use or performance of the Software, and (iii) more generally, any consequential loss. In particular the Parties expressly agree that any or all pecuniary or business loss (i.e. loss of data, loss of profits, operating loss, loss of customers or orders, opportunity cost, any disturbance to business activities) or any or all legal proceedings instituted against the Licensee by a third party, shall constitute consequential loss and shall not provide entitlement to any or all compensation from the Licensor. Article 9 - WARRANTY 9.1 The Licensee acknowledges that the scientific and technical state-of-the-art when the Software was distributed did not enable all possible uses to be tested and verified, nor for the presence of possible defects to be detected. In this respect, the Licensee's attention has been drawn to the risks associated with loading, using, modifying and/or developing and reproducing the Software which are reserved for experienced users. The Licensee shall be responsible for verifying, by any or all means, the suitability of the product for its requirements, its good working order, and for ensuring that it shall not cause damage to either persons or properties. 9.2 The Licensor hereby represents, in good faith, that it is entitled to grant all the rights over the Software (including in particular the rights set forth in Article 5). 9.3 The Licensee acknowledges that the Software is supplied "as is" by the Licensor without any other express or tacit warranty, other than that provided for in Article 9.2 and, in particular, without any warranty as to its commercial value, its secured, safe, innovative or relevant nature. Specifically, the Licensor does not warrant that the Software is free from any error, that it will operate without interruption, that it will be compatible with the Licensee's own equipment and software configuration, nor that it will meet the Licensee's requirements. 9.4 The Licensor does not either expressly or tacitly warrant that the Software does not infringe any third party intellectual property right relating to a patent, software or any other property right. Therefore, the Licensor disclaims any and all liability towards the Licensee arising out of any or all proceedings for infringement that may be instituted in respect of the use, modification and redistribution of the Software. Nevertheless, should such proceedings be instituted against the Licensee, the Licensor shall provide it with technical and legal assistance for its defense. Such technical and legal assistance shall be decided on a case-by-case basis between the relevant Licensor and the Licensee pursuant to a memorandum of understanding. The Licensor disclaims any and all liability as regards the Licensee's use of the name of the Software. No warranty is given as regards the existence of prior rights over the name of the Software or as regards the existence of a trademark. Article 10 - TERMINATION 10.1 In the event of a breach by the Licensee of its obligations hereunder, the Licensor may automatically terminate this Agreement thirty (30) days after notice has been sent to the Licensee and has remained ineffective. 10.2 A Licensee whose Agreement is terminated shall no longer be authorized to use, modify or distribute the Software. However, any licenses that it may have granted prior to termination of the Agreement shall remain valid subject to their having been granted in compliance with the terms and conditions hereof. Article 11 - MISCELLANEOUS 11.1 EXCUSABLE EVENTS Neither Party shall be liable for any or all delay, or failure to perform the Agreement, that may be attributable to an event of force majeure, an act of God or an outside cause, such as defective functioning or interruptions of the electricity or telecommunications networks, network paralysis following a virus attack, intervention by government authorities, natural disasters, water damage, earthquakes, fire, explosions, strikes and labor unrest, war, etc. 11.2 Any failure by either Party, on one or more occasions, to invoke one or more of the provisions hereof, shall under no circumstances be interpreted as being a waiver by the interested Party of its right to invoke said provision(s) subsequently. 11.3 The Agreement cancels and replaces any or all previous agreements, whether written or oral, between the Parties and having the same purpose, and constitutes the entirety of the agreement between said Parties concerning said purpose. No supplement or modification to the terms and conditions hereof shall be effective as between the Parties unless it is made in writing and signed by their duly authorized representatives. 11.4 In the event that one or more of the provisions hereof were to conflict with a current or future applicable act or legislative text, said act or legislative text shall prevail, and the Parties shall make the necessary amendments so as to comply with said act or legislative text. All other provisions shall remain effective. Similarly, invalidity of a provision of the Agreement, for any reason whatsoever, shall not cause the Agreement as a whole to be invalid. 11.5 LANGUAGE The Agreement is drafted in both French and English and both versions are deemed authentic. Article 12 - NEW VERSIONS OF THE AGREEMENT 12.1 Any person is authorized to duplicate and distribute copies of this Agreement. 12.2 So as to ensure coherence, the wording of this Agreement is protected and may only be modified by the authors of the License, who reserve the right to periodically publish updates or new versions of the Agreement, each with a separate number. These subsequent versions may address new issues encountered by Free Software. 12.3 Any Software distributed under a given version of the Agreement may only be subsequently distributed under the same version of the Agreement or a subsequent version, subject to the provisions of Article 5.3.4. Article 13 - GOVERNING LAW AND JURISDICTION 13.1 The Agreement is governed by French law. The Parties agree to endeavor to seek an amicable solution to any disagreements or disputes that may arise during the performance of the Agreement. 13.2 Failing an amicable solution within two (2) months as from their occurrence, and unless emergency proceedings are necessary, the disagreements or disputes shall be referred to the Paris Courts having jurisdiction, by the more diligent Party. Version 2.0 dated 2006-09-05. ###################################################################### CONTRAT DE LICENCE DE LOGICIEL LIBRE CeCILL Avertissement Ce contrat est une licence de logiciel libre issue d'une concertation entre ses auteurs afin que le respect de deux grands principes préside à sa rédaction: * d'une part, le respect des principes de diffusion des logiciels libres: accès au code source, droits étendus conférés aux utilisateurs, * d'autre part, la désignation d'un droit applicable, le droit français, auquel elle est conforme, tant au regard du droit de la responsabilité civile que du droit de la propriété intellectuelle et de la protection qu'il offre aux auteurs et titulaires des droits patrimoniaux sur un logiciel. Les auteurs de la licence CeCILL (pour Ce[a] C[nrs] I[nria] L[ogiciel] L[ibre]) sont: Commissariat à l'Energie Atomique - CEA, établissement public de recherche à caractère scientifique, technique et industriel, dont le siège est situé 25 rue Leblanc, immeuble Le Ponant D, 75015 Paris. Centre National de la Recherche Scientifique - CNRS, établissement public à caractère scientifique et technologique, dont le siège est situé 3 rue Michel-Ange, 75794 Paris cedex 16. Institut National de Recherche en Informatique et en Automatique - INRIA, établissement public à caractère scientifique et technologique, dont le siège est situé Domaine de Voluceau, Rocquencourt, BP 105, 78153 Le Chesnay cedex. Préambule Ce contrat est une licence de logiciel libre dont l'objectif est de conférer aux utilisateurs la liberté de modification et de redistribution du logiciel régi par cette licence dans le cadre d'un modèle de diffusion en logiciel libre. L'exercice de ces libertés est assorti de certains devoirs à la charge des utilisateurs afin de préserver ce statut au cours des redistributions ultérieures. L'accessibilité au code source et les droits de copie, de modification et de redistribution qui en découlent ont pour contrepartie de n'offrir aux utilisateurs qu'une garantie limitée et de ne faire peser sur l'auteur du logiciel, le titulaire des droits patrimoniaux et les concédants successifs qu'une responsabilité restreinte. A cet égard l'attention de l'utilisateur est attirée sur les risques associés au chargement, à l'utilisation, à la modification et/ou au développement et à la reproduction du logiciel par l'utilisateur étant donné sa spécificité de logiciel libre, qui peut le rendre complexe à manipuler et qui le réserve donc à des développeurs ou des professionnels avertis possédant des connaissances informatiques approfondies. Les utilisateurs sont donc invités à charger et tester l'adéquation du logiciel à leurs besoins dans des conditions permettant d'assurer la sécurité de leurs systèmes et/ou de leurs données et, plus généralement, à l'utiliser et l'exploiter dans les mêmes conditions de sécurité. Ce contrat peut être reproduit et diffusé librement, sous réserve de le conserver en l'état, sans ajout ni suppression de clauses. Ce contrat est susceptible de s'appliquer à tout logiciel dont le titulaire des droits patrimoniaux décide de soumettre l'exploitation aux dispositions qu'il contient. Article 1 - DEFINITIONS Dans ce contrat, les termes suivants, lorsqu'ils seront écrits avec une lettre capitale, auront la signification suivante: Contrat: désigne le présent contrat de licence, ses éventuelles versions postérieures et annexes. Logiciel: désigne le logiciel sous sa forme de Code Objet et/ou de Code Source et le cas échéant sa documentation, dans leur état au moment de l'acceptation du Contrat par le Licencié. Logiciel Initial: désigne le Logiciel sous sa forme de Code Source et éventuellement de Code Objet et le cas échéant sa documentation, dans leur état au moment de leur première diffusion sous les termes du Contrat. Logiciel Modifié: désigne le Logiciel modifié par au moins une Contribution. Code Source: désigne l'ensemble des instructions et des lignes de programme du Logiciel et auquel l'accès est nécessaire en vue de modifier le Logiciel. Code Objet: désigne les fichiers binaires issus de la compilation du Code Source. Titulaire: désigne le ou les détenteurs des droits patrimoniaux d'auteur sur le Logiciel Initial. Licencié: désigne le ou les utilisateurs du Logiciel ayant accepté le Contrat. Contributeur: désigne le Licencié auteur d'au moins une Contribution. Concédant: désigne le Titulaire ou toute personne physique ou morale distribuant le Logiciel sous le Contrat. Contribution: désigne l'ensemble des modifications, corrections, traductions, adaptations et/ou nouvelles fonctionnalités intégrées dans le Logiciel par tout Contributeur, ainsi que tout Module Interne. Module: désigne un ensemble de fichiers sources y compris leur documentation qui permet de réaliser des fonctionnalités ou services supplémentaires à ceux fournis par le Logiciel. Module Externe: désigne tout Module, non dérivé du Logiciel, tel que ce Module et le Logiciel s'exécutent dans des espaces d'adressage différents, l'un appelant l'autre au moment de leur exécution. Module Interne: désigne tout Module lié au Logiciel de telle sorte qu'ils s'exécutent dans le même espace d'adressage. GNU GPL: désigne la GNU General Public License dans sa version 2 ou toute version ultérieure, telle que publiée par Free Software Foundation Inc. Parties: désigne collectivement le Licencié et le Concédant. Ces termes s'entendent au singulier comme au pluriel. Article 2 - OBJET Le Contrat a pour objet la concession par le Concédant au Licencié d'une licence non exclusive, cessible et mondiale du Logiciel telle que définie ci-après à l'article 5 pour toute la durée de protection des droits portant sur ce Logiciel. Article 3 - ACCEPTATION 3.1 L'acceptation par le Licencié des termes du Contrat est réputée acquise du fait du premier des faits suivants: * (i) le chargement du Logiciel par tout moyen notamment par téléchargement à partir d'un serveur distant ou par chargement à partir d'un support physique; * (ii) le premier exercice par le Licencié de l'un quelconque des droits concédés par le Contrat. 3.2 Un exemplaire du Contrat, contenant notamment un avertissement relatif aux spécificités du Logiciel, à la restriction de garantie et à la limitation à un usage par des utilisateurs expérimentés a été mis à disposition du Licencié préalablement à son acceptation telle que définie à l'article 3.1 ci dessus et le Licencié reconnaît en avoir pris connaissance. Article 4 - ENTREE EN VIGUEUR ET DUREE 4.1 ENTREE EN VIGUEUR Le Contrat entre en vigueur à la date de son acceptation par le Licencié telle que définie en 3.1. 4.2 DUREE Le Contrat produira ses effets pendant toute la durée légale de protection des droits patrimoniaux portant sur le Logiciel. Article 5 - ETENDUE DES DROITS CONCEDES Le Concédant concède au Licencié, qui accepte, les droits suivants sur le Logiciel pour toutes destinations et pour la durée du Contrat dans les conditions ci-après détaillées. Par ailleurs, si le Concédant détient ou venait à détenir un ou plusieurs brevets d'invention protégeant tout ou partie des fonctionnalités du Logiciel ou de ses composants, il s'engage à ne pas opposer les éventuels droits conférés par ces brevets aux Licenciés successifs qui utiliseraient, exploiteraient ou modifieraient le Logiciel. En cas de cession de ces brevets, le Concédant s'engage à faire reprendre les obligations du présent alinéa aux cessionnaires. 5.1 DROIT D'UTILISATION Le Licencié est autorisé à utiliser le Logiciel, sans restriction quant aux domaines d'application, étant ci-après précisé que cela comporte: 1. la reproduction permanente ou provisoire du Logiciel en tout ou partie par tout moyen et sous toute forme. 2. le chargement, l'affichage, l'exécution, ou le stockage du Logiciel sur tout support. 3. la possibilité d'en observer, d'en étudier, ou d'en tester le fonctionnement afin de déterminer les idées et principes qui sont à la base de n'importe quel élément de ce Logiciel; et ceci, lorsque le Licencié effectue toute opération de chargement, d'affichage, d'exécution, de transmission ou de stockage du Logiciel qu'il est en droit d'effectuer en vertu du Contrat. 5.2 DROIT D'APPORTER DES CONTRIBUTIONS Le droit d'apporter des Contributions comporte le droit de traduire, d'adapter, d'arranger ou d'apporter toute autre modification au Logiciel et le droit de reproduire le logiciel en résultant. Le Licencié est autorisé à apporter toute Contribution au Logiciel sous réserve de mentionner, de façon explicite, son nom en tant qu'auteur de cette Contribution et la date de création de celle-ci. 5.3 DROIT DE DISTRIBUTION Le droit de distribution comporte notamment le droit de diffuser, de transmettre et de communiquer le Logiciel au public sur tout support et par tout moyen ainsi que le droit de mettre sur le marché à titre onéreux ou gratuit, un ou des exemplaires du Logiciel par tout procédé. Le Licencié est autorisé à distribuer des copies du Logiciel, modifié ou non, à des tiers dans les conditions ci-après détaillées. 5.3.1 DISTRIBUTION DU LOGICIEL SANS MODIFICATION Le Licencié est autorisé à distribuer des copies conformes du Logiciel, sous forme de Code Source ou de Code Objet, à condition que cette distribution respecte les dispositions du Contrat dans leur totalité et soit accompagnée: 1. d'un exemplaire du Contrat, 2. d'un avertissement relatif à la restriction de garantie et de responsabilité du Concédant telle que prévue aux articles 8 et 9, et que, dans le cas où seul le Code Objet du Logiciel est redistribué, le Licencié permette aux futurs Licenciés d'accéder facilement au Code Source complet du Logiciel en indiquant les modalités d'accès, étant entendu que le coût additionnel d'acquisition du Code Source ne devra pas excéder le simple coût de transfert des données. 5.3.2 DISTRIBUTION DU LOGICIEL MODIFIE Lorsque le Licencié apporte une Contribution au Logiciel, les conditions de distribution du Logiciel Modifié en résultant sont alors soumises à l'intégralité des dispositions du Contrat. Le Licencié est autorisé à distribuer le Logiciel Modifié, sous forme de code source ou de code objet, à condition que cette distribution respecte les dispositions du Contrat dans leur totalité et soit accompagnée: 1. d'un exemplaire du Contrat, 2. d'un avertissement relatif à la restriction de garantie et de responsabilité du Concédant telle que prévue aux articles 8 et 9, et que, dans le cas où seul le code objet du Logiciel Modifié est redistribué, le Licencié permette aux futurs Licenciés d'accéder facilement au code source complet du Logiciel Modifié en indiquant les modalités d'accès, étant entendu que le coût additionnel d'acquisition du code source ne devra pas excéder le simple coût de transfert des données. 5.3.3 DISTRIBUTION DES MODULES EXTERNES Lorsque le Licencié a développé un Module Externe les conditions du Contrat ne s'appliquent pas à ce Module Externe, qui peut être distribué sous un contrat de licence différent. 5.3.4 COMPATIBILITE AVEC LA LICENCE GNU GPL Le Licencié peut inclure un code soumis aux dispositions d'une des versions de la licence GNU GPL dans le Logiciel modifié ou non et distribuer l'ensemble sous les conditions de la même version de la licence GNU GPL. Le Licencié peut inclure le Logiciel modifié ou non dans un code soumis aux dispositions d'une des versions de la licence GNU GPL et distribuer l'ensemble sous les conditions de la même version de la licence GNU GPL. Article 6 - PROPRIETE INTELLECTUELLE 6.1 SUR LE LOGICIEL INITIAL Le Titulaire est détenteur des droits patrimoniaux sur le Logiciel Initial. Toute utilisation du Logiciel Initial est soumise au respect des conditions dans lesquelles le Titulaire a choisi de diffuser son oeuvre et nul autre n'a la faculté de modifier les conditions de diffusion de ce Logiciel Initial. Le Titulaire s'engage à ce que le Logiciel Initial reste au moins régi par le Contrat et ce, pour la durée visée à l'article 4.2. 6.2 SUR LES CONTRIBUTIONS Le Licencié qui a développé une Contribution est titulaire sur celle-ci des droits de propriété intellectuelle dans les conditions définies par la législation applicable. 6.3 SUR LES MODULES EXTERNES Le Licencié qui a développé un Module Externe est titulaire sur celui-ci des droits de propriété intellectuelle dans les conditions définies par la législation applicable et reste libre du choix du contrat régissant sa diffusion. 6.4 DISPOSITIONS COMMUNES Le Licencié s'engage expressément: 1. à ne pas supprimer ou modifier de quelque manière que ce soit les mentions de propriété intellectuelle apposées sur le Logiciel; 2. à reproduire à l'identique lesdites mentions de propriété intellectuelle sur les copies du Logiciel modifié ou non. Le Licencié s'engage à ne pas porter atteinte, directement ou indirectement, aux droits de propriété intellectuelle du Titulaire et/ou des Contributeurs sur le Logiciel et à prendre, le cas échéant, à l'égard de son personnel toutes les mesures nécessaires pour assurer le respect des dits droits de propriété intellectuelle du Titulaire et/ou des Contributeurs. Article 7 - SERVICES ASSOCIES 7.1 Le Contrat n'oblige en aucun cas le Concédant à la réalisation de prestations d'assistance technique ou de maintenance du Logiciel. Cependant le Concédant reste libre de proposer ce type de services. Les termes et conditions d'une telle assistance technique et/ou d'une telle maintenance seront alors déterminés dans un acte séparé. Ces actes de maintenance et/ou assistance technique n'engageront que la seule responsabilité du Concédant qui les propose. 7.2 De même, tout Concédant est libre de proposer, sous sa seule responsabilité, à ses licenciés une garantie, qui n'engagera que lui, lors de la redistribution du Logiciel et/ou du Logiciel Modifié et ce, dans les conditions qu'il souhaite. Cette garantie et les modalités financières de son application feront l'objet d'un acte séparé entre le Concédant et le Licencié. Article 8 - RESPONSABILITE 8.1 Sous réserve des dispositions de l'article 8.2, le Licencié a la faculté, sous réserve de prouver la faute du Concédant concerné, de solliciter la réparation du préjudice direct qu'il subirait du fait du Logiciel et dont il apportera la preuve. 8.2 La responsabilité du Concédant est limitée aux engagements pris en application du Contrat et ne saurait être engagée en raison notamment: (i) des dommages dus à l'inexécution, totale ou partielle, de ses obligations par le Licencié, (ii) des dommages directs ou indirects découlant de l'utilisation ou des performances du Logiciel subis par le Licencié et (iii) plus généralement d'un quelconque dommage indirect. En particulier, les Parties conviennent expressément que tout préjudice financier ou commercial (par exemple perte de données, perte de bénéfices, perte d'exploitation, perte de clientèle ou de commandes, manque à gagner, trouble commercial quelconque) ou toute action dirigée contre le Licencié par un tiers, constitue un dommage indirect et n'ouvre pas droit à réparation par le Concédant. Article 9 - GARANTIE 9.1 Le Licencié reconnaît que l'état actuel des connaissances scientifiques et techniques au moment de la mise en circulation du Logiciel ne permet pas d'en tester et d'en vérifier toutes les utilisations ni de détecter l'existence d'éventuels défauts. L'attention du Licencié a été attirée sur ce point sur les risques associés au chargement, à l'utilisation, la modification et/ou au développement et à la reproduction du Logiciel qui sont réservés à des utilisateurs avertis. Il relève de la responsabilité du Licencié de contrôler, par tous moyens, l'adéquation du produit à ses besoins, son bon fonctionnement et de s'assurer qu'il ne causera pas de dommages aux personnes et aux biens. 9.2 Le Concédant déclare de bonne foi être en droit de concéder l'ensemble des droits attachés au Logiciel (comprenant notamment les droits visés à l'article 5). 9.3 Le Licencié reconnaît que le Logiciel est fourni "en l'état" par le Concédant sans autre garantie, expresse ou tacite, que celle prévue à l'article 9.2 et notamment sans aucune garantie sur sa valeur commerciale, son caractère sécurisé, innovant ou pertinent. En particulier, le Concédant ne garantit pas que le Logiciel est exempt d'erreur, qu'il fonctionnera sans interruption, qu'il sera compatible avec l'équipement du Licencié et sa configuration logicielle ni qu'il remplira les besoins du Licencié. 9.4 Le Concédant ne garantit pas, de manière expresse ou tacite, que le Logiciel ne porte pas atteinte à un quelconque droit de propriété intellectuelle d'un tiers portant sur un brevet, un logiciel ou sur tout autre droit de propriété. Ainsi, le Concédant exclut toute garantie au profit du Licencié contre les actions en contrefaçon qui pourraient être diligentées au titre de l'utilisation, de la modification, et de la redistribution du Logiciel. Néanmoins, si de telles actions sont exercées contre le Licencié, le Concédant lui apportera son aide technique et juridique pour sa défense. Cette aide technique et juridique est déterminée au cas par cas entre le Concédant concerné et le Licencié dans le cadre d'un protocole d'accord. Le Concédant dégage toute responsabilité quant à l'utilisation de la dénomination du Logiciel par le Licencié. Aucune garantie n'est apportée quant à l'existence de droits antérieurs sur le nom du Logiciel et sur l'existence d'une marque. Article 10 - RESILIATION 10.1 En cas de manquement par le Licencié aux obligations mises à sa charge par le Contrat, le Concédant pourra résilier de plein droit le Contrat trente (30) jours après notification adressée au Licencié et restée sans effet. 10.2 Le Licencié dont le Contrat est résilié n'est plus autorisé à utiliser, modifier ou distribuer le Logiciel. Cependant, toutes les licences qu'il aura concédées antérieurement à la résiliation du Contrat resteront valides sous réserve qu'elles aient été effectuées en conformité avec le Contrat. Article 11 - DISPOSITIONS DIVERSES 11.1 CAUSE EXTERIEURE Aucune des Parties ne sera responsable d'un retard ou d'une défaillance d'exécution du Contrat qui serait dû à un cas de force majeure, un cas fortuit ou une cause extérieure, telle que, notamment, le mauvais fonctionnement ou les interruptions du réseau électrique ou de télécommunication, la paralysie du réseau liée à une attaque informatique, l'intervention des autorités gouvernementales, les catastrophes naturelles, les dégâts des eaux, les tremblements de terre, le feu, les explosions, les grèves et les conflits sociaux, l'état de guerre... 11.2 Le fait, par l'une ou l'autre des Parties, d'omettre en une ou plusieurs occasions de se prévaloir d'une ou plusieurs dispositions du Contrat, ne pourra en aucun cas impliquer renonciation par la Partie intéressée à s'en prévaloir ultérieurement. 11.3 Le Contrat annule et remplace toute convention antérieure, écrite ou orale, entre les Parties sur le même objet et constitue l'accord entier entre les Parties sur cet objet. Aucune addition ou modification aux termes du Contrat n'aura d'effet à l'égard des Parties à moins d'être faite par écrit et signée par leurs représentants dûment habilités. 11.4 Dans l'hypothèse où une ou plusieurs des dispositions du Contrat s'avèrerait contraire à une loi ou à un texte applicable, existants ou futurs, cette loi ou ce texte prévaudrait, et les Parties feraient les amendements nécessaires pour se conformer à cette loi ou à ce texte. Toutes les autres dispositions resteront en vigueur. De même, la nullité, pour quelque raison que ce soit, d'une des dispositions du Contrat ne saurait entraîner la nullité de l'ensemble du Contrat. 11.5 LANGUE Le Contrat est rédigé en langue française et en langue anglaise, ces deux versions faisant également foi. Article 12 - NOUVELLES VERSIONS DU CONTRAT 12.1 Toute personne est autorisée à copier et distribuer des copies de ce Contrat. 12.2 Afin d'en préserver la cohérence, le texte du Contrat est protégé et ne peut être modifié que par les auteurs de la licence, lesquels se réservent le droit de publier périodiquement des mises à jour ou de nouvelles versions du Contrat, qui posséderont chacune un numéro distinct. Ces versions ultérieures seront susceptibles de prendre en compte de nouvelles problématiques rencontrées par les logiciels libres. 12.3 Tout Logiciel diffusé sous une version donnée du Contrat ne pourra faire l'objet d'une diffusion ultérieure que sous la même version du Contrat ou une version postérieure, sous réserve des dispositions de l'article 5.3.4. Article 13 - LOI APPLICABLE ET COMPETENCE TERRITORIALE 13.1 Le Contrat est régi par la loi française. Les Parties conviennent de tenter de régler à l'amiable les différends ou litiges qui viendraient à se produire par suite ou à l'occasion du Contrat. 13.2 A défaut d'accord amiable dans un délai de deux (2) mois à compter de leur survenance et sauf situation relevant d'une procédure d'urgence, les différends ou litiges seront portés par la Partie la plus diligente devant les Tribunaux compétents de Paris. Version 2.0 du 2006-09-05. kissplice-2.6.2/ChangeLog000066400000000000000000000353761423273700400153010ustar00rootroot00000000000000UNRELEASED Improvements: - manpage and user_guide.pdf build process should now be reproducible 2022-04-05 Francois Gindraud * 2.6.0 -> 2.6.1 Bug fix: - Option -s 1 works again. The last version for which it worked was 2.4.0-p1 - Various warning fixes in C++ code 2022-03-16 Audric Cologne * 2.5.5 -> 2.6.0 Major bug fixes: - "--stranded" is now working correctly and should be used with stranded RNAseq libraries - "--get-mapping-info" is now working as expected kissreads update: - Kissreads is now written with C++ CMake update - conversion to "modern CMake" conventions: targets, proper scoping of CFLAGS, find_package for requirement detection. - bcalm tool is now an external dependency by default. A locally compiled version can be used as a fallback (matching previous behavior). - make install functionality has been restored. Previously broken due to bcalm submodule which installed bcalm in bin/. - userguide is built instead of using a cached pdf. Can be disabled if no latex dsitribution is present. A CI-generated pdf available online is linked in the readme. - release simplification: version number is propagated to userguide, manpage, kissplice.py frontend script. - fixes to documentation. New important release. Compared to 2.5.x, 2.6.x is: - Faster to install - Faster to run (parallelisation of kissreads) - Using information of stranded reads in kissreads (--stranded parameter) - Outputting information about read mapping on the de Bruijn Graph (--get-mapping-info) 2022-01-21 Audric Cologne * 2.5.4 -> 2.5.5 New feature: -Paralellisation of Kissreads 2021-03-05 Audric Cologne * 2.5.3 -> 2.5.4 Bug fixes: -fixed multiple definitions of variable due to new version of gcc (gcc10) 2020-06-10 Vincent Lacroix * 2.2.5 -> 2.5.3 Bug fixes: -fixed unitary tests 2020-06-04 Vincent Lacroix * 2.5.1 -> 2.5.2 Parameters: -c from 5 to 2. Modifying this parameter in version 2.5.1 was changing too much the output. Setting it back to 2 enables to capture rare variants again, for a reasonable running time cost. 2020-04-15 Vincent Lacroix * 2.5.0 -> 2.5.1 Parameters: -b from 10 to 5. Improvement of default running time at the expense of losing out complex variants. -b had been temptatively set to 10 in version 2.5.0 to enumerate more complex bubbles but this impacts running time too much. We set it back to 5. -c from 2 to 5. Improvement of default running time at the expense of losing out rare variants. 2020-03-10 Vincent Lacroix * 2.4.0-p1 -> 2.5.0 New features: -Added --stranded parameter, enables to take advantage of the strandedness of the reads, useful when genes located on different strands overlap, this is the case for instance in human cell lines infected by influenza because the virus manipulates the termination of transcription of its host, hence transcripts are much longer and possibly overlap downstream genes, causing false positive intron retention calls. -Now compatible with python3 Improvements: -Huge speed-up in graph construction, now performed using bcalm -Experimental algorithm for bubble enumeration now used by default Parameters: --mismatches from 0 to 2, as was done in Benoit-Pilven et al. --min_overlap from 3 to 5, as was done in Benoit-Pilven et al. --counts from 0 to 2, as was done in Benoit-Pilven et al. 2016-04-14 Leandro Ishi * 2.4.0 -> 2.4.0-p1 Bugs fixes: -Fixed a bug that happened if the user uses the --min_overlap parameter. 2015-10-22 Leandro Ishi * 2.3.1 -> 2.4.0 Bugs fixes: -Fixed a minor bug in which some branching bubbles were wrongly compressed in the 4-nodes compression step. This caused some rare bubbles to be missed. -Fixed a minor bug in which some linear paths remained uncompressed after the 4-nodes compression step. This caused some rare bubbles to be missed. -Fixed a bug which caused the reads in the junction AS, SB, ASSB, AB and S to be wrongly counted and another which caused a read to map to a bubble without enough overlapping bases. This may change the counts of the events and their classification whether coherent or uncoherent. -Fixed the main script not recognizing when the stack size limit was exceeded and to advise the user to execute “ulimit -s unlimited” before rerunning KisSplice. New features: -Modified the -s parameter. -s now should be always followed by an integer and it changes which types of SNPs will be output. If 0 (default), will not output SNPs. If 1, will output Type0a-SNPs. If 2, will output Type0a and Type0b SNPs. -Added a new experimental algorithm that searches for bubbles by listing all paths. The time and memory that may be used by this new algorithm is not bounded by a polynomial function on the input size, but it worked well on some instances in practice. To use this algorithm instead of the default one, it is needed to call KisSplice with parameter --experimental. -Added a memory limit to the experimental algorithm (parameter --max-memory). When using the experimental algorithm (--experimental) instead of the default one, the user may give a memory limit to this algorithm. If the algorithm uses more than this limit, it is killed. By default, this value is unlimited (i.e. no memory limit is imposed); -Added --keep-bccs parameter which is a boolean that indicates if KisSplice should keep the node/edges files for all bccs after execution (defaults to false). -Added --output-branch-count parameter which is a boolean that indicates if KisSplice should output the number of branching nodes in each path (defaults to false). -Added --keep-counts parameter which is a boolean that indicates if KisSplice should keep the .counts file after the sequencing-errors-removal step (defaults to false. We added this because the .counts file was too big sometimes and generally the user did not need to keep it (if needed, it can be recreated with -C and --keep-counts)) -Added a timestamp to the main messages of the pipeline and a summary log file on the results folder Improvements: -Improved the Sequencing error removal step (e.g.: sometimes the files output by the error removal step are already calculated, so we do not need to redo them); -Improved the way KisSplice classifies bubbles: 1) indels (Type 3) now have a shorter path length of at maximum 2k (or 2k-2, without counting the two characters of context); 2) now the maximum 10% hamming distance to classify a bubble as Type_0b considers only the variable part of the upper and lower path. -Bubbles are now output before and after read coherency module. Parameters: -m (LL_MIN) from 2k-8 to 2k-10 (not counting the two nucleotides of context. If you count them, it was from 2k-6 to 2k-8). So, more bubbles will be found now. -M (UL_MAX) from 10000 to 1000000. This enables KisSplice to find very long and rare events, and it does not impact much in runtime. --timeout now defaults to 100000s -C from 0.02 to 0.05 2014-12-17 Camille Marchet * 2.2.1 -> 2.3.0 Bugs fixed : - Reads containing only 'N': the graph construction was stopped if the file contained a read composed only of 'N's. This is was a silence bug, no error message was produced. - Problems compiling with new versions of MAC OSX (10.8+): KisSplice was not compiling with the new default C++ compiler of OSX 10.8+. Improvements : -KissReads : * speed up x2 on average * More precise control of KissReads' behaviour by decorrelating the values of k and of the seed. * -m option was changed in order to return the Id of reads that mapped on a path Parameters : --timeout default now at 10000 2014-07-24 Camille Marchet * 2.2.0 -> 2.2.1 This version fixes a bug in KissReads when a fasta/fastq file contains empty lines. Indeed KisSplice and KissReads had not the same way to treat empty lines, KisSplice part was just ignoring them while KissReads stopped the mapping at the first line of this type. As a result, many events were classified as "incoherent", because KissReads did not check on them. They are now correctly classified. It also corrects CMAKE behavior for ZLIB library. This affects only the compilation & installation of KisSplice in some cases and does not have impact on KiSplice results. 2014-06-05 Camille Marchet * 2.1.0 -> 2.2.0 Pipeline features : - New enumeration algorithm : restricted enumeration branch has been merged and former enumeration is no longer used. This new enumeration has been developped to tackle the issue of events lost in big bccs. The idea is to enumerate all bubbles with at most b branching nodes in each path, doing so we avoid repeated regions. Options and parameters : - Do not output SNPs by default. The flag (-s) outputs SNPs. - The maximum number of cycles (max_cycles) increased from 10k to 100M. - The timeout increased from 900s to 3600s. - Option -u goes back to not returning unfinished bcc by default. - -b option to set the number of branches, related to the new enumeration method(5 by default). For developpers : - smaller data for tests 2014-01-22 Camille Marchet * 2.0.0 -> 2.1.0 Pipeline features : - Bug fixed : -Error removal step erased random edges. Now fixed : discards edges according to C value. - "N" nucleotides are now ignored by the pipeline during graph building. - Now working with Minia r6025 and Minigraph r5097. - Now using 4 Bloom filters by default. Options and parameters : - New option --mismatches makes possible to allow some mismatches while KissReads' reads alignment on results. - Option -u has changed: =1 to store unfinished bcc (default) =0 to discard unfinished bcc - Default parameters have changed : k moves from 25 to 41 and M from 1000 to 10000. Outputs : - The command line is returned in the output (.o) file. - Type_0 results file now presents two types of variants : Type_0a where only one single nucleotide change is returned in the upper path Type_0b where several single nucleotide changes are returned in the upper path For developpers : - 3 Functional tests added : test each module (ks_error_removal, ks_run_module, ks_bubble_enumeration). - A test on debruijn graph construction added - It is possible to perfom profiling on KisSplice using gprof 2013-11-13 Alice Julien-Laferriere * 1.8.3 -> 2.0 -New kissreads version following developments of symbiose project: specific counts for kissplice. 4 counts are made now : ASB, AS, SB ,S, AB (for events: ASB, AB). parallel version by default -Coherence is made based on reads covering nt not on reads covering kmer. min_overlap parameters is added. A read is count only if it overlap the variable part if the events by min_overlap nt. eg: if the read overlap by min_overlap S it is count in S, by min_overlap the junction AS, etc. -New counting option for junction are therefore in kissplice.py scripts: options --counts : 0 (counts are reported as CX), 1 ( ASX, SBX, ASSBX, ABX), 2 ( ASX, SBX, SX, ASSB, ABX). See user_guide for full information. -There are k nucleotide sequences on both sides of the variations (instead of k-1 in previous version) - New options : --output-context and --output-path --output-context : if is possible, KisSplice will output the longest sequences possible before and after the events, the outputted context is in lower case --output-path: kissplice output in all_path_k{k value} file the nodes ids of the cycles -The counts are made for the SNPs with a seed of k + 1 because of the k nt context - It is now forbidden to pick an even value for k -Changes the results, they are more numerous due to distinction of switching node enable by the k nt context. Counts can be modified a little bit, because of the seed of k+1 nt for nt and the min_overlap parameters. -Efficient internal use of disk capacities. Suitable for a large range of systems from laptops to clusters. Implemented by Gunter Roeth (Bull HPC Extreme Computing ) 2013-08-02 David Parsons * 1.8.2 -> 1.8.3 New build organization: main binary kissplice remains in the bin directory, all the secondary binaries now go to lib/kissplice 2013-06-03 Vincent Miele * 1.8.1 -> 1.8.2 New kissreads version, more stable compressed reads are now valid input 2013-04-04 Alice Julien-Laferriere * 1.8.0 -> 1.8.1 Patch: it is now possible to compile and use k-mer > 31 (use cmake . -DKMERS_OVER_32=ON) -C option: default to 0.02 2013-03-28 Alice Julien-Laferriere * 1.7.1 -> 1.8.0 De Bruijn Graph construction use new Minia version (v4) much more memory efficient Dynamic error removal: a pre-processing step to remove edges with low relative coverage, which are likely sequencing errors. Uniform criteria to stop the enumeration: if a bcc reached the timeout OR the maximum number of cycles, nothing is output. This should change the results. 2012-09-01 Gustavo Sacomoto * 1.6.3 -> 1.7 Python script kissplice replaces shell script kissplice.sh 2012-04-04 Gustavo Sacomoto * 1.6.2 -> 1.6.3 Bug related to mac OS fixed 2012-03-29 Gustavo Sacomoto * 1.6 -> 1.6.2 Large k (k>32) bug fixed Proper installation using cmake / done by Vincent Miele 2012-02-14 Pierre Peterlongo * 1.5.1 -> 1.6 Possibility to chose the output directory (-o) Possibility to run kissplice from any directory Possibility to avoid the computation of SNPs and sequencing errors (-s) Updated manual 2012-02-03 Gustavo Sacomoto * 1.5 -> 1.5.1 Small bug fixed 2012-01-23 Pierre Peterongo * 1.4 -> 1.5 ranking and sorting results on read coverage and not k-mer coverage provides 5 files: uncoherent (a bit useless) Type 0: SNPS / sequencing errors Type 1: splicing events Type 2: Tandem repeats Type 3: Short indels 2012-01-17 Pierre Peterlongo * 1.2 -> 1.4 Possibility to use k larger than 32 nt (slower) or smaller than 32 (faster) 2012-01-16 Pierre Peterlongo * 1.1 -> 1.2 Better de-bruijn graph creation Read length unlimited Checks the read coherency and provides read coverage on results 2011-10-28 Gustavo Sacomoto * 1.0 -> 1.1 Fixed small bugs Improved the user guide 2011-10-24 Pierre Peterlongo * 1.0 Initial release kissplice-2.6.2/README.md000066400000000000000000000037541423273700400150010ustar00rootroot00000000000000# KisSplice Main page: http://kissplice.prabi.fr/ [User guide](https://gitlab.inria.fr/erable/kissplice/-/jobs/artifacts/master/file/prefix/share/doc/kissplice/user_guide.pdf?job=check_install_and_user_guide) from latest build. # Pre-packaged versions ## Docker You can find the latest version of KisSplice, KisSplice2RefGenome and KissDE [on Docker Hub](https://hub.docker.com/repository/docker/dwishsan/kissplice-pipeline). We also propose a [stand-alone Docker image for the KissDE Shiny App](https://hub.docker.com/repository/docker/dwishsan/kde-shiny) for KisSplice results exploration. ## Linux A package for kissplice is available on Debian / Ubuntu, but it may lag the current version. # Build from source ## Dependencies Required: - cmake >= 3.1 - C/C++11 compiler toolchain (binutils, gcc or clang) - python3 to run kissplice Recommended but optional: - [bcalm](https://github.com/GATB/bcalm) >= v2.2.2 ; as a fallback a locally compiled version of bcalm can be used by passing `-DUSE_BUNDLED_BCALM=TRUE` to cmake. A debian/ubuntu package is available. - latex toolchain with standard packages to build the user_guide ; this can be disabled by passing `-DUSER_GUIDE=OFF` to cmake ## Build Download from the latest [release](https://gitlab.inria.fr/erable/kissplice/-/releases) and uncompress it. You can also clone the git repository, but the latest code may be broken. Then compile using : ``` cd kissplice/ # Replace by the directory where the release tar.gz was uncompressed, or where the git was cloned mkdir build cd build cmake -DCMAKE_BUILD_TYPE=Release .. make -j 4 ``` After build the user guide (if not disabled) can be found in `build/doc/user_guide.pdf`. The tool can be installed to a custom directory (like `~/.local/`) with: ``` cmake -DCMAKE_INSTALL_PREFIX= .. make make install ``` # Running on the sample test ``` cd kissplice/build ./bin/kissplice -r ../sample_example/mock1.fq -r ../sample_example/mock2.fq -r ../sample_example/virus1.fq -r ../sample_example/virus2.fq ``` kissplice-2.6.2/doc/000077500000000000000000000000001423273700400142565ustar00rootroot00000000000000kissplice-2.6.2/doc/CMakeLists.txt000066400000000000000000000036541423273700400170260ustar00rootroot00000000000000option( USER_GUIDE "compile and install a pdf user_guide ; requires pdflatex and standard latex packages" ON ) if (USER_GUIDE) find_package(LATEX) if(NOT PDFLATEX_COMPILER) message(FATAL_ERROR "pdflatex not found, install latex or disable user_guide generation with -DUSER_GUIDE=OFF") endif() # add version number configure_file("${CMAKE_CURRENT_SOURCE_DIR}/user_guide.in.tex" "${CMAKE_CURRENT_BINARY_DIR}/user_guide.tex" @ONLY) # Call pdflatex twice to fix references. # Called with pwd=source_dir to have access to pictures, but -output-directory=binary_dir to prevent clutter. # -halt-on-error -interaction=nonstopmode are here to prevent any user prompt and have a clean stop+error instead. # The quiet.sh script tries to remove verbosity if VERBOSE make is not requested. add_custom_command( OUTPUT "${CMAKE_CURRENT_BINARY_DIR}/user_guide.pdf" COMMAND "${CMAKE_CURRENT_SOURCE_DIR}/quiet.sh" "${PDFLATEX_COMPILER}" -halt-on-error -interaction=nonstopmode "-output-directory=${CMAKE_CURRENT_BINARY_DIR}" "${CMAKE_CURRENT_BINARY_DIR}/user_guide.tex" COMMAND "${CMAKE_CURRENT_SOURCE_DIR}/quiet.sh" "${PDFLATEX_COMPILER}" -halt-on-error -interaction=nonstopmode "-output-directory=${CMAKE_CURRENT_BINARY_DIR}" "${CMAKE_CURRENT_BINARY_DIR}/user_guide.tex" COMMAND "${CMAKE_CURRENT_SOURCE_DIR}/rm_pdf_id.sh" "${CMAKE_CURRENT_BINARY_DIR}/user_guide.pdf" WORKING_DIRECTORY "${CMAKE_CURRENT_SOURCE_DIR}" MAIN_DEPENDENCY "${CMAKE_CURRENT_BINARY_DIR}/user_guide.tex" DEPENDS "${CMAKE_CURRENT_SOURCE_DIR}/quiet.sh" "${CMAKE_CURRENT_SOURCE_DIR}/rm_pdf_id.sh" ) add_custom_target(doc ALL DEPENDS "${CMAKE_CURRENT_BINARY_DIR}/user_guide.pdf") install(FILES "${CMAKE_CURRENT_BINARY_DIR}/user_guide.pdf" DESTINATION "${CMAKE_INSTALL_DOCDIR}") else() message(WARNING "user_guide.pdf will not be built, disabled by USER_GUIDE variable") endif() kissplice-2.6.2/doc/MBTD1_C0.png000066400000000000000000002043431423273700400161230ustar00rootroot00000000000000PNG  IHDR/ -ksBIT|dtEXtSoftwaregnome-screenshot> IDATxwtT{g{H!;.TiA(AE tQj!H2B)~k2rs#4/[III$&&2]N7ϡ JRAAAAAAAAAAAAAAAAAAAAAAAAAA,P F@BAA1QRQSiD+3uɿ1X;W*Eg 2`_;ޓ&J|RA+X!-ҢA!-Ң0h=acBVMrzD@~-IFA͚|?,>#n(M#hϥ`Xe존ɳ[~L8oXUByGAAa3@-AA! 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摳'17'|B iݏm _H* }\.۟[ ,ȧqWϟKZ0hy<,u$Q`8$J$A=8I_kaD'PE_J%mU~woʕ+W\]^;rʕ+W\})Ql۩GT-"EE;>&*{O'+gjM؆\LYwyW\nq3\rʕ+W%'gPe&'waY>sKE@/5z6c17َAqK҃4>~.vSw^M1(>ckZ#3dmYlz5CNmUჅ42zܫ{A*bsc r/.t{*ZPS)}F{TPΆ{RHƵ[(RGIFz|$ʕ|Wf4v\& BmH|un̯oHTNhLf|1ηؚj.L),;' 9At g&NٶG_Ir4__vTQBߑ:B!8$cK!9pY9oTjuuM-dSNs{Y}^ ]vtč= n=]S(zdh_T Ht| dm sl+li8:[Ԯݫ7)5Hj=VVCńX?:ǦSѶ+$ͮݾ `tB!HB!mx؛~U'_,nUB!Ŀ/!~,+B!B!'y}\&IENDB`kissplice-2.6.2/doc/user_guide.in.tex000066400000000000000000000700111423273700400175370ustar00rootroot00000000000000\documentclass[english, a4paper, 12pt]{article} \usepackage[T1]{fontenc} \usepackage[latin1]{inputenc} \usepackage{geometry} \usepackage[english]{babel} \usepackage{multicol} \usepackage{babel} \usepackage{multicol} \usepackage{babel} \usepackage{amsmath} \usepackage{amssymb} \usepackage[dvips]{graphicx} \usepackage{color} \usepackage{alltt} %\usepackage{dsfont} \usepackage{amsthm} \usepackage{enumerate} \usepackage{url} \usepackage{fancyvrb} \usepackage{listings} \lstset{ basicstyle=\small\ttfamily, columns=flexible, breaklines=true } \geometry{verbose,letterpaper,tmargin=2cm,bmargin=3cm,lmargin=2.5cm,rmargin=2.5cm} \newcommand{\soft}{\texttt{KisSplice}} \newcommand{\esoft}{\texttt{KisSplice} } \title{ \soft\\ De-novo calling alternative splicing events\\ from RNA-seq data\\ \vspace{0.5cm} User's guide, version @PROJECT_VERSION@ \vspace{-1cm} } \date{@CONFIGURE_DATE@} \author{} \begin{document} \maketitle \vspace{-1cm} \tableofcontents \newpage %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% \section{\soft, at a glance} \esoft is dedicated to {\it de-novo} calling of alternative splicing events from one or several RNA-seq datasets. In addition to splicing events, \esoft detects indels and SNPs. Data from different conditions can be co-assembled with \soft. %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% \section{\esoft software package} \subsection{CeCILL licence} This software is governed by the CeCILL licence. Details are mentioned in the \texttt{COPYING} file. \subsection{Reference} If you use \esoft in a published work, please cite the following reference:\\ Sacomoto et al. KISSPLICE: de-novo calling alternative splicing events from RNA-seq data. BMC Bioinformatics 13, S5 (2012). https://doi.org/10.1186/1471-2105-13-S6-S5\\ If you use \soft, kissplice2reftranscriptome and kissDE, please cite:\\ Lopez-Maestre et al., SNP calling from RNA-seq data without a reference genome: identification, quantification, differential analysis and impact on the protein sequence, Nucleic Acids Research, Volume 44, Issue 19, 2 November 2016, Page e148\\ If you use \soft, kissplice2refgenome and kissDE, please cite:\\ Benoit-Pilven et al. Complementarity of assembly-first and mapping-first approaches for alternative splicing annotation and differential analysis from RNAseq data. Sci Rep 8, 4307 (2018). https://doi.org/10.1038/s41598-018-21770-7 \subsection{Forum} If you have questions, please use primarily the biostars forum: \url{www.biostars.org/t/kissplice/} \subsection{Mailing list} To be informed about new releases, bugs or updates, please subscribe to the mailing list \texttt{kissplice-users@lists.gforge.inria.fr}.\\ To do so, please subscribe at : \url{http://lists.gforge.inria.fr/cgi-bin/mailman/listinfo/kissplice-users}. Do not hesitate to write to the list for any comment or questions on \soft. \subsection{Requirements} \esoft uses the \texttt{CMake} building tool (at least 3.1 CMake version is required). If it is not directly available on your system, you can download it from \url{http://www.cmake.org/cmake/resources/software.html}. \esoft relies on \texttt{zlib} that is included in most systems, however it is also available from \url{http://zlib.net}. \subsection{Installation} \esoft is written in C/C\texttt{++} and is running on Mac OS X and Linux 64 bits platforms..\\ In order to install \soft, you need to: \begin{enumerate} \item Uncompress the archive file: \begin{small} \begin{Verbatim}[frame=single] tar zxvf kissplice-x.y.z.tar.gz \end{Verbatim} \end{small} \item Go into the corresponding directory: \begin{small}\begin{Verbatim}[frame=single] cd kissplice-x.y.z \end{Verbatim} \end{small} \item Launch cmake: \begin{small} \begin{Verbatim}[frame=single] cmake . \end{Verbatim} \end{small} If \texttt{zlib} library is present in your system in an uncommon path, launch cmake with the following options: \begin{small} \begin{Verbatim}[frame=single] cmake . -DCMAKE_LIBRARY_PATH=/path/to/zlib -DCMAKE_INCLUDE_PATH=/path/to/zlib \end{Verbatim} \end{small} \item Compile the package: \begin{small} \begin{Verbatim}[frame=single] make \end{Verbatim} \end{small} \item Run KisSplice: \begin{small} \begin{Verbatim}[frame=single] ./bin kissplice \end{Verbatim} \end{small} %\item Run the functional and integration tests: %\begin{small} %\begin{Verbatim}[frame=single] %make test %\end{Verbatim} %\end{small} %\item Install the package on your system: %\begin{small} %\begin{Verbatim}[frame=single] %sudo make install %\end{Verbatim} %\end{small} \item If you want to locally install the package in a particular directory \texttt{/path\_to\_install/} (like \texttt{$\mathtt{\sim}$/.local/}), then do : \begin{small}\begin{Verbatim}[frame=single] cmake . -DCMAKE_INSTALL_PREFIX=/path_to_install/ make make install \end{Verbatim} \end{small} \end{enumerate} %If you do not install the software, the program is available in the repertory \textbf{bin}. %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% \section{Usage} \subsection{Testing \esoft in a couple of minutes} The example presented here only deals with alternative splicing events, but the format is the same for SNPs and indels. %The output format for SNPs and indels is however similar. %The example that will be presented only deals with alternative splicing events. However, other types of outputs are very similar and their interpretation can be easily inferred from this case.\\ %The \texttt{sample\_example} directory contains two files \texttt{reads1.fa} and \texttt{reads2.fa} which correspond to simulated reads from two transcripts of D. melanogaster, which differ by the inclusion of one exon. Running \esoft on this dataset should output one splicing event. The \texttt{sample\_example} directory contains four files \texttt{mock1.fq}, \texttt{mock2.fq}, \texttt{virus1.fq}, \texttt{virus2.fq} which correspond to RNAseq reads from the MBTD1 gene in two biological conditions: mock infection and infection by Influenza A virus. There are two biological replicates for each condition. Running \esoft on this dataset should output one splicing event. %Once you installed \soft, you can run it with defaults parameters (\texttt{k=41}) on the two fasta files present in the \texttt{sample\_example} directory of the release: %\begin{small} %\begin{Verbatim}[frame=single] %bin/kissplice -r sample_example/reads1.fa -r sample_example/reads2.fa %less results/results_reads1_reads2_k25_coherents_type_0.fa %\end{Verbatim} %\end{small} \begin{small} \begin{Verbatim}[frame=single] ./bin/kissplice -r sample_example/mock1.fq -r sample_example/mock2.fq -r sample_example/virus1.fq -r sample_example/virus2.fq less results/results_mock1_mock2_virus1_virus2_k41_coherents_type_1.fa \end{Verbatim} \end{small} %The result files can be found in the results directory:\\ %\texttt{results\_reads1\_reads2\_k41\_coherents\_type\_0.fa} contains the SNP/sequencing error and \texttt{results\_reads1\_reads2\_k41\_coherents\_type\_1.fa} contains the exon skipping event . %Using several \texttt{-r} can help you to co-assemble datasets. For our example, the output files can be found in the results directory:\\ \texttt{results\_mock1\_mock2\_virus1\_virus2\_k41\_coherents\_type\_1.fa} which contains the alternative splicing event (Fig. \ref{fig:extype1_C2}). The other files are empty. %In fig \ref{fig:extype0}, the content of \texttt{results\_reads1\_reads2\_k41\_coherents\_type\_0.fa}, outputting the SNPs found by \esoft. %The output is pairwise, representing the variation between two sequences. Here it is a one nucleotide change: $G \rightarrow T$ (letter in red) between the path called "upper path" and the path called "lower path". %The modification is at the center of the paths of length 49. \texttt{Type0a} indicates that there is a single SNP within the sequence (\texttt{Type0b} would indicate the presence of multiple SNPs, i.e. several SNPs separated by less than $k$ nt, or a pattern created by paralogous genes).\\ %When a SNP is indicated in the type 0 file, both paths can be merged into a single sequence, replacing the letter by an $N$. %Now, let observe the alternative splicing event reported by KisSplice (Fig. \ref{fig:extype1}). \begin{figure}[!h] \includegraphics[width=1\textwidth]{MBTD1_C2} \caption{Content of the file \texttt{results\_mock1\_mock2\_virus1\_virus2\_k41\_coherents\_type\_1.fa}, the alternative splicing event found. In red are the flanking k-mers (i.e. extremities of the flanking exons). In white is the variable part (i.e. the skipped exon).}\label{fig:extype1_C2} \end{figure} The output is organised in blocks of four lines, each corresponding to an AS event. Here there is just one block. Each block is composed of two sequence variants. The upper path corresponds to the longer variant, and the lower path to the shorter variant. The sequence of each path starts and ends with the k-mer common to both path (highlighted in red). The upper path contains an additional sequence (highlighted in white). In this example, the additional sequence is an alternatively skipped exon. The flanking k-mers are the extremities of the flanking exons. The fasta header of each path additionnally contains read counts. When using the --counts 0 option (Figure \ref{fig:extype1_C0}), the counts are easier to understand because there is just one count for each input file. \begin{figure}[!h] \includegraphics[width=1\textwidth]{MBTD1_C0} \caption{Content of the file \texttt{results\_mock1\_mock2\_virus1\_virus2\_k41\_coherents\_type\_1.fa}, when using --counts 0 option. In red are the flanking k-mers (i.e. extremities of the flanking exons). In white is the variable part (i.e. the skipped exon).}\label{fig:extype1_C0} \end{figure} In the example, the longer splice variant is supported by 47 reads in mock1, 78 reads in mock2, 18 reads in virus1, 13 reads in virus 2. The shorter splice variant is supported by 2 reads in mock1, 3 reads in mock 2, 14 reads in virus1 and 15 reads in virus2. Overall, this exon is more often skipped in samples infected with IAV. When using the --counts 2 option (default option in Kissplice-2.5.x), (Figure \ref{fig:extype1_C2}), the counts of the longer variant are split in 4 categories: reads fully contained in the variable part (S counts), reads spanning the junction with the upstream exon (AS counts), reads spanning the junction with the downstream exon (SB counts), reads spanning both junctions (ASSB counts). For this example, out of the 47 reads supporting the longer variant in mock 1: 15 support the first junction, 23 the second junction, 9 are included in the skipped exon and none support both junctions. %In the sequence identifier, we can see that the event is in the same compound than the type 0 one (the compound is here called bcc 1). %The output is pairwise, representing the variation between two sequences. Here it is an exon skipping : there is an inclusion in the path called "upper path" which is not in the path called "lower path". %There are also the counts of the two isoforms: %\begin{itemize} %\item inclusion isoform : 0 (for condition 1, reads1.fa file) , 50 (for condition 2, reads2.fa file) %\item exclusion isoform : 7 , 0 %\end{itemize} %\begin{figure}[!h] %\includegraphics[width=1\textwidth]{alt_splicing_sample_ex} %\caption{Content of the file \texttt{results\_reads1\_reads2\_k41\_coherents\_type\_1.fa}, the alternative splicing event found. In red is the variable part in the inclusion isoform.}\label{fig:extype1} %\end{figure} %The splicing event is condition-specific with the inclusion isoform covered by 50 reads in condition 2 and the exclusion isoform covered by 7 reads in condition 1.\\ % You may want to map the sequences against a reference genome if you have one to see what the output corresponds to. This can be done using blat, STAR, HiSat2 or any other RNA/DNA alignment software. This step can be automatised using %\begin{figure} %\includegraphics[width=1\textwidth]{ucsc.png} %\caption{Using the reference genome, it is possible to see that the type 1 event is an exon skipping (figure obtained using Blat and the UCSC genome browser)} %\end{figure} % \newpage \subsection{Common TroubleShooting} Before running \esoft on a large dataset, (10 conditions with 100M reads each), which takes time and memory, we advise to first run \esoft on a subset of your dataset (2 conditions with 10M reads each) and get familiar with the output. If you encounter problems running \soft, this is often due to the stack size limit. Please try increasing your stack size before running \soft. The best way to do this is setting the stack size to unlimited, if your operating system accepts this, executing the following command: \begin{small} \begin{Verbatim}[frame=single] > ulimit -s unlimited \end{Verbatim} \end{small} If your operating system does not accept the previous command, you can increase your stack size to the highest value possible. To find this value, execute: \begin{small} \begin{Verbatim}[frame=single] > ulimit -H -s \end{Verbatim} \end{small} And then, to set the stack size, execute: \begin{small} \begin{Verbatim}[frame=single] > ulimit -s \end{Verbatim} \end{small} If the problem persists, please do not hesitate to contact us. \subsection{Options} Type \texttt{kissplice -h} to see the full list of options and parameters \subsection{Counts option } %\esoft can use several methods to quantify the paths found in the DBG. %The quantification depends on the \texttt{min\_overlap} parameters. A read is used for quantification if and only if it overlaps (by at least \texttt{min\_overlap} nt) the variable part of the path. We can distinguish various parts in the paths of an event (please see Fig. \ref{fig:quantif}): \begin{itemize} \item AS: junction between the left part of the path (A in green) and the variable part of the longer path (S in red). A read is in AS if it has more than \texttt{min\_overlap} nt in S and A \item SB: junction between the right part of the path (B in blue), and the variable part of the longer path (S in red). A read is in SB if it has more than \texttt{min\_overlap} nt in S and B. \item S: the variable part (in red), the difference between the upper and lower path. A read is in S if it has less than \texttt{min\_overlap} nt in A or B. \item AB: junction between A and B (left, resp. right part of the path) in the shorter path. A read is in AB if it has more than \texttt{min\_overlap} nt in A and B. \item ASSB: the junction AS and SB with S. A read is counted in ASSB if if has more than \texttt{min\_overlap} nt in A and B (and so if fully in S). \end{itemize} \begin{figure}[h!] \includegraphics[width=1\textwidth]{quantifModel.png} \caption{Example of quantification. In lower case, the context obtained using the \texttt{--output-context} parameter.} \label{fig:quantif} \end{figure} The \texttt{--counts} options can take three values: \texttt{0,1,2}. If does not affect the quantification itself, but changes the output. By default, in kissplice-2.5.x, the counts option is set to 2. This affects only splicing events. \begin{enumerate}[start=0] %\setcounter{enumi}{0} \item the total count is reported (default behaviour) as CX \item the counts are written for ASX, SBX, ASSBX, ABX \item all the counts are reported (ASX, SBX, SX, ASSBX and ABX) \end{enumerate} Here X indicates the read file used for the quantification. It is possible to retrieve the total count C using: $C = (AS-ASSB) + (SB-ASSB) + S + ASSB$ \subsection{Paired-end reads} For now, no particular treatment is given to paired-end reads. We recommend that you use them as two input files : \begin{small} \begin{Verbatim}[frame=single] kissplice -r condition1/1 -r condition1/2 \end{Verbatim} \end{small} \section{I/O} \subsection{Input} \esoft may be used either directly from one or several sets of reads, or from a bi-directed de-Bruijn graph.\\ In the first case, one or several fasta/fastq files containing the reads have to be provided. In the second case, the de-Bruijn graph alone has to be provided (in dot format). In the input fasta/q files, each read should be written on exactly two lines for fasta (one for the fasta identifier and one for the sequence) and four lines for fastq. \\ The input files should have one of the following extensions: \texttt{fq, fastq, txt} or \texttt{fasta, fa}. \esoft also handles compressed reads (\texttt{.fa.gz}) To have an example of the input files, run \esoft on the test data (provided in the directory \texttt{sample\_example}), consisting of two read files in fasta format. You will find the constructed de-Bruijn graph built by \esoft in the \esoft directory. \subsection{Output}\label{subsec:output} If it was not provided, a de-Bruijn graph is built for the $N$ fasta files (read) given as input; the following three files are created in the results directory (for $k=XX$): \begin{itemize} \item \texttt{graph\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX.nodes} for the nodes of the de-Bruijn graph \item \texttt{graph\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX.edges} for its edges. %\item \texttt{graph\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX.solid\_kmers\_binary\_with\_count} a binary file relevant for the option \texttt{-C } %\item \texttt{graph\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX.counts} for the $k-mers$ count, created only if \esoft was run with option \texttt{-C} coupled with \texttt{-\-keep-counts}. \item \texttt{graph\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX.abundance} for the $k-mers$ count of each node. \end{itemize} In the node file, the 1st column is the node ID, the 2nd its (forward) sequence. In the file describing the edges, the 1st and 2nd column are the IDs of the nodes that are connected by an edge, the 3rd column codes for the direction of the edge (FF = from the forward sequence of a node to the forward sequence of the other node, RR= from reverse to reverse, FR and RF). Note that if node A is connected with node B by an edge with direction ``FF'' (``FR''), node B is connected to node A by an edge directed ``RR'' (``FR'') and vice versa. Moreover, six fasta files for the results are created: \begin{enumerate} \item \texttt{results\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX\_type\_0a.fa}: Single SNPs or sequencing substitution errors \item \texttt{results\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX\_type\_0b.fa}: Multiple SNPs or sequencing substitution errors \item \texttt{results\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX\_type\_1.fa}: alternative splicing events \item \texttt{results\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX\_type\_2.fa}: inexact tandem repeats \item \texttt{results\_NameReadFile1\_NameReadFile2\_...\_NameReadFileN\_kXX\_type\_3.fa}: short indels \item \texttt{results\_NameReadFile1\_...\_NameReadFileN\_kXX\_type\_4 .fa}: all others, composed by a shorter path of length $> 2k$ not being a SNP \end{enumerate} If the read files (option \texttt{-r}) are provided as input files, the fasta files are checked for read coherency, which means that each nucleotide of each sequence has to be covered by at least one read. If \esoft is used with option \texttt{-g} (only the de-bruijn graph is provided), a check for read coherency is not possible due to missing information about read coverage. \begin{sloppypar} Since the read coherency step may take a long time, \esoft preemptively outputs these six fasta files before checking for read coherency in the directory \texttt{results\_without\_read\_coherency} inside the results directory. This enables the user to access the results even before the read coherency module finishes. \esoft also creates a summary log file in the results directory named \texttt{kissplice\_log\_summary\_(time)\_(date)\_(random\_integer)} in case the user wants to view a summary of the execution. This is not meant to be a detailed output. If you need a detailed output, please run \esoft in verbose mode (parameter \texttt{-v}) and redirect the output and error stream to a file. \end{sloppypar} Fasta files are organized as follows, each 4-lines groups are an event \begin{small} \begin{Verbatim}[frame=single] > identifier_upper_path sequence upper path > identifier_lower_path sequence lower path \end{Verbatim} \end{small} The identifier for the upper path is formatted as follows : \begin{small} \begin{Verbatim}[frame=single] >bcc_BB|Cycle_YY|Type_ZZ|upper_path_Length_UU|C1_cov1|C2_cov2[...]|CN_covN|rank_RR \end{Verbatim} \end{small} \begin{itemize} \item \texttt{bcc\_BB} : Bi-connected component BB the event belongs to \item \texttt{Cycle\_YY} : since in each bcc, several cycles may exist, this attribute indicates the ID of the cycle (here: cycle YY) that generated the bubble \item \texttt{Type\_ZZ}: with \texttt{ZZ=\{0a, 0b, 1, 2, 3, 4\}}, the type also corresponds to the type given by the file name \item \texttt{upper\_path\_Length\_UU} : length (in nucleotides) of the sequence of the upper path of the bubble \item \texttt{Cn}: with \texttt{n=\{1,..., N\}}: coverage of the path using reads from the read file n; coverage is the raw count of reads mapping the path with at least \texttt{k} (i.e. the value specified for the $k-mer$ length) nucleotides. \item \texttt{rank\_RR} : %The rank is close to 1 if the alleles of the event are condition-specific. Otherwise, it is close to 0. Formally, the rank corresponds to the square of the Phi coefficient. In the case where there is one condition, the rank is always 0. In the case where there are more than two conditions, it is calculated for all pairs of conditions, and the maximal phi value is output. Note that the rank is just an indication of the strength of the association between the allele and a condition but this is not a statistical test. In particular, for low counts, the rank may be high but a statistical test may give a negative answer to the question ``is there an association between alleles and conditions ?''. On the other hand, for very large counts, a statistical test may give a positive answer while the effect is very small. The rank helps to measure the strength of the effect. This piece of information used to give an indication of if the alleles/the variants were differentially expressed between two conditions. It was an indication of the strength of the association between the allele/the variant and a condition but this was not a statistical test. We now provide a reliable indicator based on statistical analysis in our package kissDE (see \url{http://kissplice.prabi.fr/tools/kissDE/}). \end{itemize} The identifier for the lower path provides virtually the same information, but concerning the lower (shorter) path of the bubble. In order to facilitate the further post-processing of these files, the bcc, cycle, type and rank are repeated, although this information is redundant. If \esoft is run with option \texttt{-r} (read files), the events in the output files are sorted with respect to their rank (this is not possible with option -g due to missing information about read coverage). \subsection{Uncoherent Bubbles} In order for a bubble to be considered as coherent, each nt has to be covered by at least one read. Uncoherent bubbles are output in a separate file. In principle, uncoherent bubbles should correspond to artefacts of the DBG (we lose information when we move from reads to $k-mers$). In practice, real events which have a low coverage may produce uncoherent bubbles. Hence, it may be worth to mine this file if you are interested in unfrequent events. \section{Performances} \subsection{Pre-treatment} \esoft can be run on raw data. However, as any assembler, it performs better if the data is pre-treated. Common pre-treatment includes polyA tail removal and quality filter. These filters are not included in \esoft distribution but can easily be found. For instance, FASTX Toolkit can be used to trim the reads, i.e. removing the last bases for which quality is below a threshold (we commonly use 20). Reads shorter than 20 nt are then removed. \subsection{Job calibration} The first time you run \esoft, you may be interested in getting quickly initial results. For moderate size datasets (<500M reads), this can be achieved with default parameters. For large datasets (>500M reads), this may require to modify some parameters. By default $c$ is set to 1. This means that only $k$-mers seen more than once are considered for the analysis. $k$-mers seen only once are indeed likely to correspond to sequencing errors. They can also correspond to rare variants. Increasing $c$ to 5 will correspond to a major speed-up of \esoft, at the expense of a loss in sensitivity: variants supported by less than 5 reads will not be reported anymore. By default $k$ is set to 41. This means that repeats smaller than 41nt will not cause trouble. This also means that reads have to overlap by at least 40nt to be assembled. Increasing $k$ to 51 will enable to solve more complex regions due to repeats, and hence will speed-up \esoft, at the expense of a loss in sensitivity: reads will need to overlap by at least 50nt to be assembled. You might be wondering about the disk space you need to have available for a job. \esoft writes in several files and temporary files, and we currently consider that you must have two to three times the size of your data in free space to run \esoft under proper conditions. To resolve disk space issues, we recommand to use \texttt{-d} option followed by a directory able to contain between two and three times the size of the input data.\\ If you want to use parallelisation option in your job, our advice is to put \texttt{-t} at 4 or 6 processors, more is not really efficient. \subsection{Relative coverage} The option \texttt{-C} edits the De-Bruijn graph constructed by removing edges non covered. It is a local filter. For a specific node, if one of its outgoing edges is covered less than \texttt{C} (in percentage) it is removed. For a small value of \texttt{C}, this option allows to remove artificial edges due to overlapping of two $k-mers$ (not supported by the reads) or sequencing errors. Increasing \texttt{-C} will reduce the run time as there will be less things to enumerate, but the price is to lose some events with a rare isoform. \subsection{Big datasets} %For some datasets, enumerating all bubbles is very long. Hence, we set a maximum amount of time (900s by default) that the algorithm spends in each BCC (biconnected component). If after this time, the BCC is not finished, the algorithm stops and moves to the next BCC. In practice, these unfinished BCCs correspond to very repetitive regions of the dataset (transposable elements...). For some datasets, enumerating all bubbles is very long. Hence, we set a maximum amount of time (100000s by default) that the algorithm spends in each BCC (biconnected component). If after this time, the BCC is not finished, the algorithm stops and moves to the next BCC. Running \esoft with \texttt{-u} option enables to output these unfinished BCCs for further inspection (for instance using Cytoscape).\\ If you intend to work on rather big datasets (> 1G reads), have a look at your \texttt{.e} log file in the end of the run. If you read "\texttt{Timeout reached}", it means the maximum amount of time has been reached, some bccs may have been lost then. We recommand to run again KisSplice with a higher \texttt{-C} (\texttt{-C} 0.05 is a good input) to simplify the graph (while not losing too much information as there are already many reads) and retrieve all events, or to increase the timeout (option \texttt{--timeout}). %\subsection{Genome-size option} %The \texttt{-z} option is used for a rough estimation of the graph size. It is an estimation of unique $k-mers$ contained into the dataset. %A good evaluation of this parameter leads to an optimal graph construction step in term of memory consumption and time. %On one hand, if the value is too low, the construction will take fewer memory but will take more time. On the other hand, if the value is too high, it will use a lot of memory. %By default \texttt{z} is set to a billion (\texttt{1 000 000 000}) which is a good trade-off for most datasets. \section{Further questions} Please visit our at FAQ \url{http://kissplice.prabi.fr/FAQ/}. %\bibliographystyle{unsrt} %\bibliography{} \end{document} kissplice-2.6.2/kissplice.in.py000077500000000000000000002277041423273700400164750ustar00rootroot00000000000000#!/usr/bin/env python3 # *************************************************************************** # # KisSplice # de-novo calling alternative splicing events from RNA-seq data. # # *************************************************************************** # # Copyright INRIA # contributors : Vincent Lacroix # Pierre Peterlongo # Gustavo Sacomoto # Alice Julien-Laferriere # David Parsons # Vincent Miele # Leandro Lima # Audric Cologne # # pierre.peterlongo@inria.fr # vincent.lacroix@univ-lyon1.fr # # This software is a computer program whose purpose is to detect alternative # splicing events from RNA-seq data. # # This software is governed by the CeCILL license under French law and # abiding by the rules of distribution of free software. You can use, # modify and/ or redistribute the software under the terms of the CeCILL # license as circulated by CEA, CNRS and INRIA at the following URL # "http://www.cecill.info". # As a counterpart to the access to the source code and rights to copy, # modify and redistribute granted by the license, users are provided only # with a limited warranty and the software's author, the holder of the # economic rights, and the successive licensors have only limited # liability. # In this respect, the user's attention is drawn to the risks associated # with loading, using, modifying and/or developing or reproducing the # software by the user in light of its specific status of free software, # that may mean that it is complicated to manipulate, and that also # therefore means that it is reserved for developers and experienced # professionals having in-depth computer knowledge. Users are therefore # encouraged to load and test the software's suitability as regards their # requirements in conditions enabling the security of their systems and/or # data to be ensured and, more generally, to use and operate it in the # same conditions as regards security. # # The fact that you are presently reading this means that you have had # knowledge of the CeCILL license and that you accept its terms. import os import re import sys import time import shlex import struct import shutil import os.path import tempfile import argparse import threading import multiprocessing from random import randint from operator import itemgetter from subprocess import Popen, PIPE, STDOUT TIMEOUT=100000 MAXTIMEOUT=(2 ** 63 / 10 ** 9) - 1 # see https://stackoverflow.com/questions/45704243/what-is-the-value-of-c-pytime-t-in-python?rq=1 logFile = 0 logFileName = "" unfinished_bccs = [] num_snps = {} ############### NEW GLOBAL VALUES -> REDUNDANCE ############### # Maybe switch some of them to parameters? ENTROPYMAX=1.8 # If an upper path have less than this entropy, its cycle is deleted MAXLENGTHIDIOTIC=200 # Do idiotic strategie for upper paths of the same length and length >= this value MAXLENGTHALIGN=1000 # Do not try to align upper paths with a length >= this value (will miss some rare cases redundancy) BASEDIFFPATH_UP=2 # Base max difference of length between two upper path for them to be compared (try redundancy removal) BASEDIFFPATH_LOW=5 # Base max difference of length between two lower path for them to be compared (try redundancy removal) BASEMM_LOW=3 # Base max edit distance (levenshtein) allowed between 2 lower paths BASEMM_UP=3 # Base max edit distance (levenshtein) allowed between 2 upper paths WILDECARDS=["N","I"] # Levenshtein distance parameters EXT=1 # Malus for gap extension MATCH=0 # Malus for match MMATCH=1 # Malus for mismatch GAP=1 # Malus for gap openning # Entropy WLEN=41 # Window length on sequence to compute entropy WSLIDE=41 # Right slide of the window on the sequence ############################################################### ############### NEW FUNCTIONS -> ENTROPY ############### # Natural log aproximation using ln(x)=lim(x->inf) n(x**(1/n)-1) def ln(x): n = 10000.0 # Increase this number for more accurate estimation return n * ((x ** (1/n)) - 1) LN2=ln(2) # For conversion between natural and base 2 log def entropyShannon(s): n=0.0 dN={} # d[nucleotide]=occurence for e in s: n+=1 if e not in dN.keys(): dN[e]=0 dN[e]+=1 entShan=0 for base in dN.keys(): pN=dN[base]/n entShan+=pN*(ln(pN)/LN2) return entShan*-1 def windowEntropy(s, wLen, wSlide): # Compute Shannon entropy on a sequence s for each window of length wLen with a slide of length wSlide. lE=[] # list of entropy start=0 # start index n=len(s) for start in list(range(n))[::wSlide]: end=start+wLen if end >= n: start=n-wLen end=n-1 if start<0: start=0 lE.append(entropyShannon(s[start:end])) return lE ########################################################## ############### NEW FUNCTIONS -> REDUNDANCE ############### def idioticConsensus(s1,s2,maxEdit=3): # Make consensus base per base (s1 and s2 have the same length) cons="" nN=0 #print(s1) #print(s2) for i in range(len(s1)): if s1[i]==s2[i]: cons+=s1[i] else: cons+="N" if "N" not in [s1[i],s2[i]]: nN+=1 i+=1 if not i%100 and nN>maxEdit: #print(1) #print(cons) return [None,None] if nN>maxEdit: #print(2) #print(cons) return [None,None] #print("OK") #print(cons) return [nN,cons] def consensus(s1,s2,mTB, trimN=True): # 2 sequences and a traceback matrix # Return one of the possible consensus, prioritazing match/mmatch, then T then L # Create consensus sequence between 2 seq, adding "n" for indel and "N" for mismatches # Will strip "N" of the consensus of trimN=True nR=len(mTB)-1 nC=len(mTB[0])-1 cons="" prev="D" # previous type of alignment t=mTB[nR][nC] while t!="E": if prev in t: # If possible, continue the same type of alignment do=prev elif "D" in t: # else, priority to D do="D" elif "T" in t: do="T" else: do="L" if do=="D": if s1[nC-1]==s2[nR-1]: cons+=s1[nC-1] elif s1[nC-1]=="I" or s2[nR-1]=="I": cons+="I" else: cons+="N" nR-=1 nC-=1 else: cons+="I" if do=="T": nR-=1 else: nC-=1 prev=do t=mTB[nR][nC] cons=cons[::-1] if trimN: cons=cons.strip("IN") return cons def rc(s): # reverse complement an ATGCN sequence d={"A":"T","T":"A","G":"C","C":"G","N":"N","I":"I"} return "".join([d[e] for e in s[::-1]]) def levDist(a,b, match=MATCH, mm=MMATCH, gap=GAP, ext=1, semiLoc=False, minRC=True, maxEdit=3, doConsensus=True, trimN=True): # Levenshtein distance between two strings a and b of length n and m. Option for Semi-Local method (smallest sequence must fully aligng anywhere on the largest sequence) # Semi-Local: To do this, no initialisation of the first row (insertion are cost-free before aligning the first base of the smallest sequence) and cost-free insertion on the last row (insertion are cost-free after aligning the last base of the smallest sequence) # a is the largest sequence (switched if needed in the function) # ext : gap extension cost # minRC : also do reverse complement computation and take the minimum distance of the two # maxEdit : stop process if the final score will be greater or equal to this number. -1 to keep everything. # doConsensus : compute consensus sequence # Maybe add a maximum indel length allowed, but this could be complicated... # 'N' is considered as a wildcard cons=None extD=ext extI=ext n=len(a) m=len(b) if n=maxEdit: minLev=None add="" if minLev==T: add+="T" if minLev==L: add+="L" if minLev==D: add+="D" if add!="": mTB[r][c]=add mLev[r][c]=minLev c+=1 r+=1 if allNone: # If the whole line was made of None values # Then the alignment failed r=m+1 # Exit the loop lev=mLev[m][n] if doConsensus and lev!=None: cons=consensus(a,b,mTB,trimN=trimN) #print(mLev) #print(mTB) isRC=False if minRC and lev==None: lLevRC=levDist(a,rc(b),match=match, mm=mm, gap=gap, ext=ext, maxEdit=maxEdit, minRC=False,semiLoc=semiLoc,trimN=trimN) levRC=lLevRC[0] if levRC!=None and (lev==None or levRCmaxID): return [{},lComp] dLev={} j=0 while j+1 < nID: ID1=lID[j] l1l=d[ID1][1][1] l1u=d[ID1][1][0] ID2=lID[(j+1)] l2l=d[ID2][1][1] l2u=d[ID2][1][0] levL=None levU=None addErrorMin=int(l2u/300) addErrorMax=int(l1u/100) #print("Comparing "+ID1+" and "+ID2) if (ID1,ID2) not in lExclude and abs(l1l-l2l)<=BASEDIFFPATH_LOW and abs(l1u-l2u)<=BASEDIFFPATH_UP+addErrorMin: # Length difference of lower path < 6 and length diff of upper path < 3 # LOW COMPARISON # We had the variable sequence to the middle of the lower path var1=d[ID1][0][2] var2=d[ID2][0][2] seqLow1=d[ID1][0][1] #seqLow1=seqLow1[:k]+var1+seqLow1[k:] seqLow2=d[ID2][0][1] #seqLow2=seqLow2[:k]+var2+seqLow2[k:] levL,isRC,consL=levDist(seqLow1,seqLow2,maxEdit=BASEMM_LOW,ext=EXT,semiLoc=True) # We allow an high error rate on the lower path if levL!=None: # UP COMPARISON seqUp1=d[ID1][0][0] seqUp2=d[ID2][0][0] if isRC: # if reverse complement was used for lower path, use it for upper path seqUp2=rc(seqUp2) var2=rc(var2) # Variable path comp lLevV=levDist(var1,var2,mm=MMATCH/(addErrorMax+1),ext=EXT,semiLoc=False,minRC=False,trimN=False) levV=lLevV[0] consVar=lLevV[2] if levV!=None: # Variable path align correctly ## Test if we are in a big IR cluster idioticOK=False if len(seqUp1)==len(seqUp2) and l1u>=MAXLENGTHIDIOTIC: # Both path of the same length and upper path > 2000nc, probably a big IR levU,consU=idioticConsensus(seqUp1,seqUp2,maxEdit=BASEMM_UP+addErrorMax-levV) if levU!=None: idioticOK=True ## if not idioticOK and l1ubcc_"+bcc+"|Cycle_"+c,seqUp,">lower",seqLow])) if not bcc in dBCC2lc.keys(): dBCC2lc[bcc]=[] dBCC2lc[bcc].append([c,str(lEnt)]) return "KO" # SeqUp will be the upper sequence without the potential repeated bases at the begining/end of the upperpath, ie : # >bcc_9988|Cycle_0|Type_1|upper_path_length_163 # GGCTGCAACCGAGTCTTCATAGAAGAGAATCTGCTGTACCTCGGAATCCTCGCTGAAGTCTTCGGTGACGGTAGAGGAGGAGGCCTGCCGGGGGAGCTTGGCCTCGTATGCCATGACGCTCCACCTGTCCAGCATCTTGGTGCTGGCTCTCTCCAACTTCTCC # >bcc_9988|Cycle_0|Type_1|lower_path_length_78 # GGCTGCAACCGAGTCTTCATAGAAGAGAATCTGCTGTACCTGTCCAGCATCTTGGTGCTGGCTCTCTCCAACTTCTCC # In this exemple, the upper sequence can either be : # CGGAATCCTCGCTGAAGTCTTCGGTGACGGTAGAGGAGGAGGCCTGCCGGGGGAGCTTGGCCTCGTATGCCATGACGCTCCACCT # OR # ACCTCGGAATCCTCGCTGAAGTCTTCGGTGACGGTAGAGGAGGAGGCCTGCCGGGGGAGCTTGGCCTCGTATGCCATGACGCTCC # Because of the starting/ending ACCT # We will report seqUp=CGGAATCCTCGCTGAAGTCTTCGGTGACGGTAGAGGAGGAGGCCTGCCGGGGGAGCTTGGCCTCGTATGCCATGACGCTCC and var=ACCT # The size of var is 2*k-lowerPathLength # In can happen that the upper path is < 2*k, in this case we have to use the lvar=lup-llow lvar=mk-llow var=seqUp[k-lvar:k] seqUp=seqUp[k:k+lup-lvar] # It is possible that seqUp is empty, if the path of the var is the same as the upper path, ie # >bcc_9962|Cycle_0|Type_1|upper_path_length_82 # ATAAAGGATATGTTGAATACACCTTTGTGTCCTTCACACAGCAGTTTACATCCAGTGCTGTTACCTTCAGATGTATTTGACC # >bcc_9962|Cycle_0|Type_1|lower_path_length_79 # ATAAAGGATATGTTGAATACACCTTTGTGTCCTTCACACAGTTTACATCCAGTGCTGTTACCTTCAGATGTATTTGACC # In this exemple, seqUp='' and var="CAG" # So we simply invert them if seqUp=="": seqUp,var=var,seqUp return [ [bcc, c, t, lup, llow], [seqUp, seqLow, var]] def compress(dLev, dSeq, lCycleOrder, lExclude): # Remove one of the paths from dSeq and lCycleOrder # Return dictionnary of compressed paths #print(dLev) #print("BEFORE COMPRESS") #print(dSeq) #print(lCycleOrder) for ID1 in dLev.keys(): for ID2 in dLev[ID1].keys(): dSeq[ID1][0][0]=dLev[ID1][ID2][3] dSeq[ID1][0][2]=dLev[ID1][ID2][4] dSeq[ID1][0][1]=dLev[ID1][ID2][2] dSeq[ID1][1][0]=len(dSeq[ID1][0][0])+len(dSeq[ID1][0][2]) dSeq[ID1][1][1]=len(dSeq[ID1][0][1]) del dSeq[ID2] del lCycleOrder[lCycleOrder.index(ID2)] # Delete pairs containing either ID1 or ID2 from lExclude lExclude=[lExclude[i] for i in range(len(lExclude)) if ID1 not in lExclude[i] and ID2 not in lExclude[i]] del dLev #print("AFTER COMPRESS") #print(dSeq) #print(lCycleOrder) def addComp(d,l): # Add compressed paths to the dictionnary of compressed path # d : d[cycle]=[compCycle1, compCycle2, ...] # l : [(cycle1, cycleComp1), (cycle2, cycleComp2), ...] cycleCompX is compressed in cycleX # cycleCompX can be a key of d for cycle,comp in l: if cycle in d.keys(): d[cycle].append(comp) else: d[cycle]=[comp] if comp in d.keys(): d[cycle].extend(d[comp]) del d[comp] def compressBcc(dBcc, cBcc, dSeq, lCycleOrder, dBcc2cycleComp, dBCC2size, dBccLen2nCompress, k): # t is the type of event # Compressed sequences will be written in fComp, an open writable file # Do the whole compression of a BCC lExclude=[] # list of cycle pairs that can not be compressed # dBccLen2nCompress is not mendatory, contain useful(?) informations. d[BCCsize]=[nBCC, number of compressed path] dBcc2cycleComp[cBcc]={} # not mendatory, contain useful(?) informations. d[bcc][cycle]=[compressed cycles] BCCsize=len(dSeq.keys()) dBCC2size[cBcc]=BCCsize # not mendatory, contain useful(?) informations. d[bcc]=size if not BCCsize in dBccLen2nCompress.keys(): dBccLen2nCompress[BCCsize]=[0,0] dBccLen2nCompress[BCCsize][0]+=1 #print("BCC "+cBcc+" (of "+str(len(dSeq.keys()))+" cycles)") dBcc[cBcc],lComp=multLev(dSeq,lCycleOrder,lExclude=lExclude,k=k) while dBcc[cBcc] != {}: # While we have some compression to do #print(lComp) dBccLen2nCompress[BCCsize][1]+=len(lComp) addComp(dBcc2cycleComp[cBcc],lComp) compress(dBcc[cBcc], dSeq, lCycleOrder, lExclude) dBcc[cBcc],lComp=multLev(dSeq,lCycleOrder,lExclude=lExclude,k=k) #print(dBcc[cBcc]) #print(dBccLen2nCompress) #print(dBcc2cycleComp) def writeCompressedCycles(dSeq, cBcc, t, fComp, kval): # Write info in dSeq to fComp # dSeq: d[cycle]=[ [seqUp, seqLow, var], [lenUp, lenLow] ] # head format: >bcc_[cBcc]|Cycle_[cycle]|Type_[t]|upper/lower_path_length_[length] headBcc=">bcc_"+cBcc headType="Type_"+t for cycle in dSeq.keys(): lInfo=dSeq[cycle] seqLow=lInfo[0][1] seqUp=seqLow[:kval]+lInfo[0][0]+lInfo[0][2]+seqLow[kval:] lenUp=str(lInfo[1][0]+lInfo[1][1]) lenLow=str(lInfo[1][1]) headCycle="Cycle_"+cycle headLenUp="upper_path_length_"+lenUp headLenLow="lower_path_length_"+lenLow head="|".join([headBcc,headCycle,headType]) headUp="|".join([head,headLenUp]) headLow="|".join([head,headLenLow]) fComp.write("\n".join([headUp,seqUp,headLow,seqLow])+"\n") def splitT1T234(fName, fNameT1, fNameT234): f=open(fName,"r") f1=open(fNameT1,"w") f234=open(fNameT234,"w") retype = re.compile('Type_\d+') line=f.readline() while line: t=retype.search(line).group() if t=="Type_1": oF=f1 else: oF=f234 oF.write(line) line=f.readline() oF.write(line) line=f.readline() oF.write(line) line=f.readline() oF.write(line) line=f.readline() f.close() f1.close() f234.close() def redundancyAndLowComplexityRemoval(workdir, mainFileName, keep_rd=False, keep_lc=False, lc_ent=ENTROPYMAX, get_rd_info=True, get_lc_info=True, t1o=False, kval=41): # Main function for redundancy and low-complexity bubbles removal # workdir: str, working directory # mainFileName: str, fasta file name containing all types of bubbles # keep_rd: boolean, do we remove redundancy? # keep_lc: boolean, do we keep low-complexity bubbles? # lc_ent: int, Shannon Entropy threshold to define a bubble as low-complexity (if below this value) # get_rd_info: boolean, do we print useful(?) informations about redundancy removal in some files? # get_lc_info: boolean, do we print useful(?) informations about low-complexity removal in some files? # kval: int, k-mers value # return list of files to copy from the workdir to the resultdir, will replace mainFile by a new file if needed, and will create new files in the workdir # Do we need to do anything? if keep_rd and keep_lc and not t1o: return [] print("\n" + getTimestamp() + "--> Removing low-complexity/redundant bubbles...") printlg("\n" + getTimestamp() + "--> Removing low-complexity/redundant bubbles...") toMove=[] # list of files to move to the result directory (will be returned) toRm=[] # list of files to remove # 1) Divide Type_1 bubbles in one file, other bubbles in another file # The Type_1 file will be moved to the result directory t1fileName="all_bcc_type1.fa" t234fileName="all_bcc_type234.fa" if not keep_lc or not keep_rd: toMove.append(t1fileName) toRm.append(t234fileName) splitT1T234("/".join([workdir,mainFileName]), "/".join([workdir,t1fileName]), "/".join([workdir,t234fileName])) if not keep_rd or not keep_lc: # 2) Define some dictionnaries... dSeq={} # d[cycle]=[ [seqUp, seqLow, var], [lenUp, lenLow] ] dBcc={} # d[bcc]=dLev with dLev = d[cycle1][cycle2]=[levDistLow,levDistUp,consensusLow,consensusUpSeq,consensusUpVar] dBcc2cycleComp={} # d[bcc][cycle]=[compressed cycles] dBccLen2nCompress={} # d[BCCsize]=[nBCC, number of compressed path] dBCC2size={} # d[bcc]=size dBCC2lc={} # d[bcc]=[ [removed cycle due to low complexity, entropy value], ...] # ... and an output file t1fileNameComp="all_bcc_type1_compressed.fa" fComp=open("/".join([workdir,t1fileNameComp]), "w") if not keep_rd: toMove.append(t1fileNameComp) # 3) Open and read first line of Type_1 file f=open("/".join([workdir,t1fileName]),"r") lFasta="KO" while lFasta and lFasta=="KO": lFasta=readFasta4(f, k=kval, rmEntropy=not keep_lc, entropy_threshold=lc_ent, dBCC2lc=dBCC2lc) # [ [bcc, cycle, type, length_up, length_low], [seq_up, seq_low, seq_var] ] # Fasta file not empty and the cycle was not removed by entropy filter (!=KO) if lFasta: # First interesting line of fasta cBcc=lFasta[0][0] # current BCC dSeq[lFasta[0][1]]=[lFasta[1], lFasta[0][3:5]] # we add the sequences informations associated to this cycle to dSeq lCycleOrder=[lFasta[0][1]] # list of cycles ID order as in the fasta file (close cycles have less divergence) # 4) Read the whole fasta file and compress BCC while lFasta: # Read a new cycle lFasta=readFasta4(f, k=kval, rmEntropy=not keep_lc, entropy_threshold=lc_ent, dBCC2lc=dBCC2lc) # [ [bcc, cycle, type, length_up, length_low], [seq_up, seq_low, seq_var] ] if lFasta and lFasta!="KO": # EOF or removed cycle due to low complexity if cBcc!=lFasta[0][0]: # New bcc # Compress the cycles from the previous BCC, if needed if not keep_rd: compressBcc(dBcc, cBcc, dSeq, lCycleOrder, dBcc2cycleComp, dBCC2size, dBccLen2nCompress, kval) writeCompressedCycles(dSeq, cBcc, "1", fComp, kval) cBcc=lFasta[0][0] # New current BCC dSeq={} lCycleOrder=[] dSeq[lFasta[0][1]]=[lFasta[1], lFasta[0][3:5]] # we add the sequences informations associated to this cycle to dSeq lCycleOrder.extend([lFasta[0][1]]) # list of cycles ID order as in the fasta file (close cycles have less divergence) # Compress the cycles of the last BCC if not keep_rd: compressBcc(dBcc, cBcc, dSeq, lCycleOrder, dBcc2cycleComp, dBCC2size, dBccLen2nCompress, kval) writeCompressedCycles(dSeq, cBcc, "1", fComp, kval) f.close() fComp.close() # 5) Make informations files if get_rd_info: fNameSummary="get_redundancy_info_summary.tsv" fNameRd="get_redundancy_info_compressed_bubbles.tsv" toMove.extend([fNameSummary,fNameRd]) makeSummaryRd("/".join([workdir,fNameSummary]), "/".join([workdir,fNameRd]), dBcc2cycleComp, dBccLen2nCompress) if get_lc_info: fNameLc="get_low-complexity_info.tsv" toMove.append(fNameLc) makeSummaryLc("/".join([workdir,fNameLc]), dBCC2lc) # 6) Write a new mainFile, combining filtered type 1 and type 234 or type 1 only if t1o: if not keep_rd or not keep_lc: os.system("cat "+"/".join([workdir,t1fileNameComp])+" > "+"/".join([workdir,mainFileName])) else: os.system("cat "+"/".join([workdir,t1fileName])+" > "+"/".join([workdir,mainFileName])) else: os.system("cat "+"/".join([workdir,t1fileNameComp])+" "+"/".join([workdir,t234fileName])+" > "+"/".join([workdir,mainFileName])) # 7) Remove some files for fRm in toRm: os.system("rm "+"/".join([workdir,fRm])) print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") return toMove def makeSummaryLc(fNameLc, dBCC2lc): # dBCC2lc : d[bcc]=[ [removed cycle due to low complexity, entropy value], ...] fLc=open(fNameLc, "w") # low-complexity file # bcc removed_cycle entropy_value head="\t".join(["bcc", "removed_cycle", "shannon_entropy"]) fLc.write(head) for bcc in dBCC2lc.keys(): for lInfo in dBCC2lc[bcc]: rmCycle=lInfo[0] ent=str(lInfo[1]) fLc.write("\n"+"\t".join([bcc, rmCycle, ent])) fLc.close() def makeSummaryRd(fNameSummary, fNameRd, dBcc2cycleComp, dBccLen2nCompress): # dBcc2cycleComp : d[bcc][cycle]=[compressed cycles] # dBccLen2nCompress : d[BCCsize]=[nBCC, number of compressed path] fSum=open(fNameSummary, "w") fRd=open(fNameRd, "w") # redundancy file # bcc consensus_cycle compressed_cycles nCompressed headRd="\t".join(["bcc","consensus_cycle","compressed_cycles","nCompressed"]) fRd.write(headRd) for bcc in dBcc2cycleComp.keys(): for cycle in dBcc2cycleComp[bcc].keys(): lComp=dBcc2cycleComp[bcc][cycle] if lComp!=[]: fRd.write("\n"+"\t".join([bcc, cycle, ",".join(lComp), str(len(lComp))])) fRd.close() # summary file # bcc_size nBcc nCycles nCompressedCycles %compressed nRemainingCycles headSum="\t".join(["bcc_size","nBcc","nCycles","nCompressedCycles", "%compressed", "nRemainingCycles"]) fSum.write(headSum) for bccSize in sorted(list(dBccLen2nCompress.keys())): lInfo=dBccLen2nCompress[bccSize] nBcc=lInfo[0] nComp=lInfo[1] nCycles=bccSize*nBcc nRemain=nCycles-nComp pComp=round((nComp/nCycles)*100) fSum.write("\n"+"\t".join([str(bccSize), str(nBcc), str(nCycles), str(nComp), str(pComp)+"%", str(nRemain)])) fSum.close() ########################################################### # print str to the logFile def printlg (*args): global logFile print(''.join(str(arg) for arg in args), file=logFile) # get the timestamp as string def getTimestamp(): return "["+time.strftime("%H:%M:%S")+" "+time.strftime("%d/%m/%Y")+"] " class Command(object): # deprecated in the future with Python3 def __init__(self): self.process = None def target(self, **kwargs): self.process = Popen(kwargs["args"], stdout=kwargs["stdout"], stderr=PIPE) com = self.process.communicate() if com[0] and (kwargs["verbose"] or self.process.returncode != 0): print(com[0]) # Prints stderr that was piped by Popen if com[1] and (kwargs["verbose"] or self.process.returncode != 0): print(com[1]) def run(self, command_line, out_file = "", mode = 'w', verbose = False, timeout = MAXTIMEOUT): if verbose: print(getTimestamp() + "Running "+command_line) args = shlex.split(command_line) if len(out_file): stdout_file = open(out_file, mode) kwargs = {"verbose":verbose, "args":args, "stdout":stdout_file} else: kwargs = {"verbose":verbose, "args":args, "stdout":PIPE} # Create a Thread object that will run "self.target" with arguments kwargs # (given in the form of keyword argument) and start it thread = threading.Thread(target=self.target, kwargs=kwargs) thread.start() # Wait for end of thread or time out thread.join( timeout ) # Check whether thread has ended or timed out # (if timed out, kill it and wait for it to actually die) if thread.is_alive(): self.process.terminate() thread.join() if len(out_file): stdout_file.close() if self.process.returncode == -15: print("\n\t\t *** Timeout reached! ***\n", file=sys.stderr) #+ command_line elif self.process.returncode == 15: print("\n\t\t *** Maximum number of bubbles reached! ***\n", file=sys.stderr) elif self.process.returncode == -6: print("\n\t\t *** Memory limit reached! ***\n", file=sys.stderr) elif self.process.returncode == -11: print("\n\t\t *** Problem with " + command_line.split()[0] + " ***", file=sys.stderr) print("\t\t *** Try increasing your stack size before running KisSplice executing: \"ulimit -s unlimited\" (if your OS accepts it, otherwise, you can replace \"unlimited\" by the value returned when executing \"ulimit -H -s\").***\n", file=sys.stderr) sys.exit(self.process.returncode) elif self.process.returncode != 0: print("Problem with " + command_line.split()[0], file=sys.stderr) sys.exit(self.process.returncode) def mkdirTmp(tmpdir=None): if not tmpdir: workdir = tempfile.mkdtemp(prefix="kissplice.") else: workdir = tempfile.mkdtemp(prefix="kissplice.", dir=tmpdir) return workdir def cleanTmp(workdir): shutil.rmtree(workdir) def subprocessLauncher(command_line, out_file = "", mode = 'w', verbose = False, timeout = MAXTIMEOUT): command = Command() command.run(command_line, out_file, mode, verbose, timeout) return command.process.returncode def to_file(readfiles, filename = "tmp"): f = open(filename, 'w') reads = readfiles.split(' ') for r in reads: f.write(os.path.abspath(r) + "\n") f.close() #BCALM to KS graph format def BCALMUnitigs2DotNodes(inputFileName, outputFileName): with open(inputFileName) as BCALMFile, open(outputFileName, "w") as dotNodesFile: for line in BCALMFile: line=line.strip() if line.startswith(">"): dotNodesFile.write(line.split()[0][1:]) #print just the id of the node else: dotNodesFile.write("\t%s\n"%line) def BCALMStrand2KSStrand(strand): if strand=="-": return "R" elif strand=="+": return "F" else: dieToFatalError("Error on BCALMStrand2KSStrand(). Offending strand: %s"%strand) #BCALM to KS edges format def BCALMUnitigs2DotEdges(inputFileName, outputFileName): allEdges=[] with open(inputFileName) as BCALMFile, open(outputFileName, "w") as dotEdgesFile: for line in BCALMFile: line=line.strip() if line.startswith(">"): words = line.split() sourceNodeId = int(words[0][1:]) for word in words: if word.startswith("L:"): fields = word.split(":") sourceNodeStrand = BCALMStrand2KSStrand(fields[1]) targetNodeId = int(fields[2]) targetNodeStrand = BCALMStrand2KSStrand(fields[3]) allEdges.append((sourceNodeId, targetNodeId, sourceNodeStrand+targetNodeStrand)) allEdges.sort() for edge in allEdges: dotEdgesFile.write("%d\t%d\t%s\n"%(edge[0], edge[1], edge[2])) #BCALM to KS unitigs abundance format def BCALMUnitigs2DotAbundance(inputFileName, outputFileName): with open(inputFileName) as BCALMFile, open(outputFileName, "w") as dotAbundanceFile: for line in BCALMFile: line=line.strip() if line.startswith(">"): words = line.split() for word in words: if word.startswith("km:f:"): fields = word.split(":") dotAbundanceFile.write("%s\n"%(fields[-1])) # Run debruijn graph construction def build_graph(internal_bindir, workdir, readfiles, kval, graphfile, min_cov, nbCores, verbose = False): print(getTimestamp() + "--> Building de Bruijn graph...") printlg(getTimestamp() + "--> Building de Bruijn graph...") print("Graph will be written in "+graphfile+".[edges/nodes]") printlg("Graph will be written in "+graphfile+".[edges/nodes]") #put all readfiles in a file all_read_files = workdir + "/all_read_filenames" to_file(readfiles, all_read_files) all_read_files = os.path.abspath(all_read_files) #to execute BCALM, we change wd because it produces the files in the wd if not os.path.exists(workdir+"/bcalm"): os.makedirs(workdir+"/bcalm") previousWD = os.getcwd() os.chdir(workdir+"/bcalm") #execute BCALM command_line = "%s/bcalm -in %s -kmer-size %d -abundance-min %d -nb-cores %d -out bcalm_out"%(internal_bindir, all_read_files, kval, min_cov, nbCores) subprocessLauncher(command_line, verbose=verbose) os.chdir(previousWD) #transform BCALM unitigs 2 ks unitigs BCALMUnitigs2DotNodes(workdir+"/bcalm/bcalm_out.unitigs.fa", graphfile + ".nodes") #build .edges file from BCALM unitigs file BCALMUnitigs2DotEdges(workdir+"/bcalm/bcalm_out.unitigs.fa", graphfile + ".edges") #build the unitigs count file from BCALM unitigs file BCALMUnitigs2DotAbundance(workdir+"/bcalm/bcalm_out.unitigs.fa", graphfile + ".abundance") print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") #Run error_removal for the graph (overwrite edge file) def error_removal(internal_bindir, graphfile, nobuild, cutoff, verbose = False): print("\n" + getTimestamp() + "--> Removing sequencing errors...") printlg("\n" + getTimestamp() + "--> Removing sequencing errors...") #checks if user passed the graph for KisSplice. In this case, maybe the error_removal step is already done if nobuild: #checks if the file created by the error_removal step is already done edge_suffix = "_C"+str(cutoff)+".edges" if os.path.isfile(graphfile+edge_suffix): print("Sequencing-errors-removal step skipped: using previously computed file "+graphfile+edge_suffix) printlg("Sequencing-errors-removal step skipped: using previously computed file "+graphfile+edge_suffix) print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") return True #Run the error-removal command_line = internal_bindir+"/ks_error_removal "+graphfile+".edges "+graphfile+".abundance "+str(cutoff)+" "+graphfile+"_C"+str(cutoff) subprocessLauncher(command_line, verbose=verbose) print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") return True #Run the modules on the graph def run_modules(internal_bindir, workdir, graphfile, kval, cutoff, verbose = False, output_context = False, exec_error_removal = False): if not os.path.exists(workdir+"/bcc"): os.mkdir(workdir+"/bcc") print("\n" + getTimestamp() + "--> Finding the BCCs...") printlg("\n" + getTimestamp() + "--> Finding the BCCs...") edge_suffix = ".edges" if exec_error_removal: edge_suffix = "_C"+str(cutoff)+".edges" command_line = internal_bindir+"/ks_run_modules "+graphfile+edge_suffix+" "+graphfile+".nodes "+str(kval)+" "+workdir+"/bcc/graph" if output_context: command_line += " --output-context" return_code = subprocessLauncher(command_line, verbose=verbose) if (return_code != 0): print("\t\t *** Try increasing your stack size before running KisSplice executing: \"ulimit -s unlimited\" (if your OS accepts it, otherwise, you can replace \"unlimited\" by the value returned when executing \"ulimit -H -s\").***\n") print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") def count_alreadyfoundSNPs(workdir): global num_snps info_snp_file = open(workdir+"/bcc/graph_info_snp_bcc", 'r') info_snp = info_snp_file.readlines() for bcc_snp in info_snp: info = bcc_snp.split()# format: bcc_id num_snps num_snps[info[0]] = int(info[1]) info_snp_file.close() def find_bcc_ids_ordered_by_size(workdir, min_length = 4): f = open( workdir+"/bcc/graph_info_bcc") bccnum2size = f.readlines()[2:] bccnumorderedbysize = [int(e[0])+1 for e in sorted(enumerate([int(t.split()[1]) for t in bccnum2size]), key=lambda x:x[1], reverse=True) if int(e[1]) >= min_length ] f.close() return (bccnum2size, bccnumorderedbysize) def enumerate_all_bubbles(internal_bindir, workdir, outdir, kval, bval, output_snps, min_edit_dist, max_cycles, UL_MAX, LL_MAX, LL_MIN, timeout, nbprocs=1, verbose = False, output_context = False, output_path = False, output_branch_count = False, experimental = False, max_memory = 0): print("\n" + getTimestamp() + "--> Enumerating all bubbles...") printlg("\n" + getTimestamp() + "--> Enumerating all bubbles...") if os.path.isfile(workdir+"/all_bcc_type0_"+str(kval)): os.remove(workdir+"/all_bcc_type0_"+str(kval)) if os.path.isfile(workdir+"/all_bcc_type1234_"+str(kval)): os.remove(workdir+"/all_bcc_type1234_"+str(kval)) f = open(workdir+"/bcc/graph_info_bcc") n_bcc = int(f.readline()) f.close() file2size = {} # filling num_snps count_alreadyfoundSNPs(workdir); # ordering bcc by decreasing size and filtering if <4 nodes bccnum2size, bccnumorderedbysize = find_bcc_ids_ordered_by_size(workdir, 4) if verbose: if len(bccnumorderedbysize) != len(bccnum2size): print("Less than 4 nodes, cannot contain a bubble!") # multiprocessing step- BEGIN pool = multiprocessing.Pool(nbprocs) TASKS = [] for i in bccnumorderedbysize: TASKS += [(enumerate_bubbles_core, i, internal_bindir, workdir, outdir, kval, bval, output_snps, min_edit_dist, max_cycles, UL_MAX, LL_MAX, LL_MIN, timeout, verbose, output_context, output_path, output_branch_count, experimental, max_memory)] imap_unordered_it = pool.imap_unordered(eval_func_tuple, TASKS, 1) for x in imap_unordered_it: if x != -1: unfinished_bccs.append(x) pool.close() pool.join() # multiprocessing step - END destinationSNPS = open(workdir+"/all_bcc_type0_"+str(kval), 'wb') ## THE FILE CONTAINS SNPS destination1234 = open(workdir+"/all_bcc_type1234_"+str(kval), 'wb') ## THE FILE CONTAINS other bcc for file in os.listdir(workdir): if file[0:17] == "tmp_all_bcc_type0": shutil.copyfileobj(open(workdir+"/"+file, 'rb'), destinationSNPS) if file[0:20] == "tmp_all_bcc_type1234": shutil.copyfileobj(open(workdir+"/"+file, 'rb'), destination1234) destinationSNPS.close() destination1234.close() if output_path: destination_paths = open(workdir+"/all_paths_k"+str(kval), 'wb') for file in os.listdir(workdir): if file[0:18] == "tmp_all_paths_bcc_": shutil.copyfileobj(open(workdir+"/"+file, 'rb'), destination_paths) destination_paths.close() f = open(workdir+"/all_bcc_type0_"+str(kval)) size0 = sum(1 for line in f) f.close() f = open(workdir+"/all_bcc_type1234_"+str(kval)) size1234 = sum(1 for line in f) f.close() n_bubbles = (size0 + size1234)/4 print("Total number of bubbles found: ", n_bubbles) printlg("Total number of bubbles found: ", n_bubbles) print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") def enumerate_bubbles_core(i, internal_bindir, workdir, outdir, kval, bval, output_snps, min_edit_dist, max_cycles, UL_MAX, LL_MAX, LL_MIN, timeout, verbose = False, output_context = False, output_path = False, output_branch_count = False, experimental = False, max_memory = 0): if verbose: print("@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@") print("Enumerating bubbles in biconnected component "+str(i)) infofile = workdir+"/bcc/graph_info_bcc" contents_file_edges = workdir+"/bcc/graph_contents_edges_bcc" contents_file_nodes = workdir+"/bcc/graph_contents_nodes_bcc" basename_edges = workdir+"/bcc/graph_all_edges_bcc" basename_nodes = workdir+"/bcc/graph_all_nodes_bcc" # Contains -1 if the process finished or the bcc number if it timed out. flag = -1 # already num_snps found - it is also the starting number from enumerating cycle num_snps_bcc = 0 if str(i) in num_snps: num_snps_bcc = num_snps[str(i)] command_line = internal_bindir+"/ks_bubble_enumeration "+ infofile+" "+ contents_file_edges+" "+ contents_file_nodes+" "+ basename_edges+" "+ basename_nodes\ +" "+str(i) \ +" "+str(kval)+" "+workdir+"/bcc/tmp_bcc_sequences_"+str(kval)+"_"+multiprocessing.current_process().name+" "+str(min_edit_dist) \ +" bcc_"+str(i) + " " + str(num_snps_bcc) + " -u "+str(UL_MAX) \ +" -L "+str(LL_MAX)+" -l "+str(LL_MIN)+" -M "+str(max_cycles)+" -s "+str(output_snps) if output_context: command_line += " -c" if output_path: command_line += " -p" if bval is not None: command_line += " -b" + str(bval) if output_branch_count: command_line += " -v" if experimental: command_line += " -e " + str(max_memory) process_returncode = subprocessLauncher(command_line, verbose=verbose, timeout=timeout) # Store the bcc number if it timed out (return code -15) OR the maximum number of bubbles was reached (return code 15) OR the memory limit was exceeded (return code -6) if process_returncode == -15 or process_returncode == 15 or process_returncode == -6: flag = i # Always append the results if the enumeration, even when it's incomplete. command_line_type0 = internal_bindir+"/ks_clean_duplicates " + workdir + "/bcc/tmp_bcc_sequences_" + str(kval) +"_"+multiprocessing.current_process().name+ "_type0.fa" command_line_type1234 = internal_bindir+"/ks_clean_duplicates " + workdir + "/bcc/tmp_bcc_sequences_" + str(kval) +"_"+multiprocessing.current_process().name+ "_type1234.fa" subprocessLauncher(command_line_type0, workdir+"/tmp_all_bcc_type0_"+str(kval)+"_"+multiprocessing.current_process().name, 'a', verbose=verbose) # append ALL BCC IN THE SAME FILE subprocessLauncher(command_line_type1234, workdir+"/tmp_all_bcc_type1234_"+str(kval)+"_"+multiprocessing.current_process().name, 'a', verbose=verbose) # append ALL BCC IN THE SAME FILE if output_path: command_line = "cat "+workdir+"/bcc/tmp_bcc_sequences_" + str(kval) +"_"+multiprocessing.current_process().name+ ".path" subprocessLauncher(command_line, workdir+"/tmp_all_paths_bcc_"+str(kval)+"_"+multiprocessing.current_process().name, 'a', verbose=verbose) # append ALL BCC IN THE SAME FILE if verbose: print("@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@\n") return flag def eval_func_tuple(f_args): return f_args[0](*f_args[1:]) def concatenate_graph_all_log_bcc_to_all_bcc_type0(workdir, kval, output_snps): if output_snps==2: #concatenate all non-branching snps destinationSNPS = open(workdir+"/all_bcc_type0_"+str(kval), 'a') ## THE UNIQUE FILE ALSO CONTAINS SNPS shutil.copyfileobj(open(workdir+"/bcc/graph_all_log_bcc", 'r'), destinationSNPS) destinationSNPS.close() elif output_snps==1: #concatenate only non-branching Type-0a destinationSNPS = open(workdir+"/all_bcc_type0_"+str(kval), 'a') ## THE UNIQUE FILE ALSO CONTAINS SNPS #append the Type_0a bubbles to the destinationSNPS file snpsFile = open(workdir+"/bcc/graph_all_log_bcc", 'r') writeLine = False for line in snpsFile.readlines(): if writeLine == True: destinationSNPS.write(line) writeLine = False else: if ("Type_0a" in line): destinationSNPS.write(line) writeLine = True else: writeLine = False destinationSNPS.close() def check_read_coverage_and_sort_all_bubbles(internal_bindir, readfiles, workdir, outdir, kval, output_snps, infix_name, countsMethods, minOverlap, substitutions, substitutionsSNP, getMappingInfo, stranded, strandedAbsoluteThreshold, strandedRelativeThreshold, nbprocs, verbose = False): # Two KisSreads executions, one for type 0 one for type 1234 # Du to the k extension, anchor should be of size k+1 for SNP commandLineType0="" if output_snps > 0: commandLineType0 = internal_bindir+"/ks_kissreadsSNPS "+workdir+"/all_bcc_type0_"+str(kval)+" "+readfiles+" -i 5 -S 25 -O "+str(kval+minOverlap)+" -o "+ workdir+"/coherentType0.fa -u "+workdir+"/uncoherentType0.fa -d " + str(substitutionsSNP) + " -c 1 -n -t "+str(nbprocs) if stranded: commandLineType0+=" -x -a " + str(strandedAbsoluteThreshold) + " -r " + str(strandedRelativeThreshold) + " " if getMappingInfo: commandLineType0+=" -m " + workdir+"/mapping_info_reads_on_Type0_bubbles.sam " subprocessLauncher(commandLineType0, verbose=verbose) #move the mapping info file if getMappingInfo: shutil.move(workdir+"/mapping_info_reads_on_Type0_bubbles.sam", outdir+"/mapping_info_reads_on_Type0_bubbles.sam") #move the explanation file if os.path.exists(workdir+"/uncoherentType0.fa.explanations"): shutil.move(workdir+"/uncoherentType0.fa.explanations", outdir+"/uncoherentType0.fa.explanations") print("\n" + getTimestamp() + "--> Computing read coherence and coverage...") printlg("\n" + getTimestamp() + "--> Computing read coherence and coverage...") # no n options anymore commandLineType1234 = internal_bindir+"/ks_kissreadsSplice "+workdir+"/all_bcc_type1234_"+str(kval)+" "+readfiles+" -i 5 -k "+str(kval)+" -S 25 -O "+str(kval+minOverlap)+" -o "+workdir+"/coherentType1234.fa -u "+workdir+"/uncoherentType1234.fa -d " + str(substitutions) + " -c 1 -j " + countsMethods +" -l " + str(minOverlap) +" -t "+str(nbprocs) if stranded: commandLineType1234+=" -x -a " + str(strandedAbsoluteThreshold) + " -r " + str(strandedRelativeThreshold) + " " if getMappingInfo: commandLineType1234+=" -m " + workdir+"/mapping_info_reads_on_Type1234_bubbles.sam" subprocessLauncher(commandLineType1234, verbose=verbose) if getMappingInfo: shutil.move(workdir+"/mapping_info_reads_on_Type1234_bubbles.sam", outdir+"/mapping_info_reads_on_Type1234_bubbles.sam") #move the explanation file if os.path.exists(workdir+"/uncoherentType1234.fa.explanations"): shutil.move(workdir+"/uncoherentType1234.fa.explanations", outdir+"/uncoherentType1234.fa.explanations") commandLineCat = "cat " + workdir+"/uncoherentType1234.fa " if output_snps > 0: commandLineCat += workdir+"/uncoherentType0.fa " subprocessLauncher(commandLineCat, workdir + "/uncoherent.fa", "a", verbose=verbose ) print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") print(getTimestamp() +"--> Sorting all bubbles...") printlg(getTimestamp() +"--> Sorting all bubbles...") nb = [0]*6# counter of number of events of each type found eventsName = ["type_0a", "type_0b", "type_1", "type_2", "type_3", "type_4"] cofilel = [] for i in range(0,6): cofilel.append(open(outdir+"/results_"+infix_name+"_coherents_"+eventsName[i]+".fa", 'w')) if output_snps > 0: snpsFile = open(workdir+"/coherentType0.fa", 'r') l = snpsFile.readlines() l.sort( reverse = True ) snpsFile.close() for event in l: eventSplitted = event.split()[-1].replace(';','\n') try: if ("Type_0a" in eventSplitted): cofilel[0].write(eventSplitted+"\n")#Transform to Fasta type nb[0] += 1 else: cofilel[1].write(eventSplitted+"\n")#Transform to Fasta type nb[1] += 1 except: pass # handling coherent "other" cofile = open(workdir+"/coherentType1234.fa", 'r') l = cofile.readlines() l.sort(reverse=True) cofile.close() retype = re.compile('Type_\d+') for event in l: try: type = retype.search(event).group() for i in range(2,6): if (type=="Type_"+str(i-1)): cofilel[i].write(event.split()[-1].replace(';','\n')+"\n")#Transform to Fasta type nb[i] += 1 except: pass for i in range(0,6): cofilel[i].close() uncofile = open(workdir+"/uncoherent.fa", 'r') uncofileout = open(outdir+"/results_"+infix_name+"_uncoherent.fa", 'w') for event in uncofile.readlines(): uncofileout.write(event.split()[-1].replace(';','\n')+"\n") uncofile.close() uncofileout.close() print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") return nb def sort_all_bubbles(internal_bindir, readfiles, workdir, outdir, kval, output_snps, infix_name, shouldDoReadCoherence, verbose = False): print("\n" + getTimestamp() + "--> Starting Bubble Output Module") printlg("\n" + getTimestamp() + "--> Starting Bubble Output Module") if shouldDoReadCoherence: outdir = outdir+"/results_without_read_coherency" if not os.path.exists(outdir): os.mkdir(outdir) print("Before checking for read coherency, all bubbles will be written to folder " + outdir) printlg("Before checking for read coherency, all bubbles will be written to folder " + outdir) print("This enables you to access them even before the read coherency module finishes, which can take a long time") printlg("This enables you to access them even before the read coherency module finishes, which can take a long time") print(getTimestamp() + "--> Sampling bubbles by type...") printlg(getTimestamp() + "--> Sampling bubbles by type...") concatenate_graph_all_log_bcc_to_all_bcc_type0(workdir, kval, output_snps) retype = re.compile('Type_\d+') eventsName = ["type_0a", "type_0b", "type_1", "type_2", "type_3", "type_4"] filel = [] for i in range(0,6): filel.append(open(outdir+"/results_"+infix_name+"_"+eventsName[i]+".fa", 'w')) nb = [0]*6 if output_snps > 0: snpsFile = open(workdir+"/all_bcc_type0_"+str(kval), 'r') for line in snpsFile.readlines(): if "Type_0a" in line: ofile = filel[0] nb[0] += 1 elif "Type_0b" in line: ofile = filel[1] nb[1] += 1 ofile.write(line) snpsFile.close() # handling the other type cfile = open(workdir+"/all_bcc_type1234_"+str(kval), 'r') for line in cfile.readlines(): try: type = retype.search(line).group() for i in range(1,5): if (type=="Type_"+str(i)): ofile = filel[i+1] nb[i+1] += 1 except: pass ofile.write(line) cfile.close() for i in range(0,6): nb[i] /= 2 filel[i].close() print(getTimestamp() + "--> Done!") printlg(getTimestamp() + "--> Done!") print("You can now access all bubbles without read coherency in: " + outdir) printlg("You can now access all bubbles without read coherency in: " + outdir) return nb def save_bccs_from_list(bcc_list, dir_name, internal_bindir, workdir, outdir, verbose = False): if not os.path.exists(outdir + dir_name): os.mkdir(outdir + dir_name) infofile = workdir+"/bcc/graph_info_bcc" contents_file_edges = workdir+"/bcc/graph_contents_edges_bcc" contents_file_nodes = workdir+"/bcc/graph_contents_nodes_bcc" basename_edges = workdir+"/bcc/graph_all_edges_bcc" basename_nodes = workdir+"/bcc/graph_all_nodes_bcc" for i in bcc_list: command_line = internal_bindir+"/ks_print_bcc "+ infofile+" "+ contents_file_edges+" "+ contents_file_nodes+" "+ basename_edges+" "+ basename_nodes\ +" "+str(i)+" "\ + outdir+ dir_name + "/graph_bcc_"+str(i)+".edges "\ + outdir+ dir_name + "/graph_bcc_"+str(i)+".nodes" subprocessLauncher(command_line, verbose=verbose) def check_read_files(readfiles): if readfiles is None: return True allFilesAreOK = True for file in readfiles: if not os.path.isfile(file): print("[ERROR] File \""+file+"\" does not exist.") allFilesAreOK = False if not allFilesAreOK: dieToFatalError("One or more read files do not exist.") def dieToFatalError (msg): print("[FATAL ERROR] " + msg) print("Try `kissplice --help` for more information") global logFileName os.remove(logFileName) sys.exit(1) # ############################################################################ # Main # ############################################################################ def main(): # script_bin_dir : absolute path to the main executable (this file), computed at runtime # @KISSPLICE_BINDIR_TO_INTERNAL_BINDIR@ : relative path from main script to internal binaries, set by cmake # internal_bindir : absolute path to the secondary executables (eg ks_kissreads) script_bindir = os.path.dirname(os.path.abspath(sys.argv[0])) internal_bindir = os.path.realpath(os.path.join(script_bindir, '@KISSPLICE_BINDIR_TO_INTERNAL_BINDIR@')) # ======================================================================== # Manage command line arguments # ======================================================================== parser = argparse.ArgumentParser(description='kisSplice - local assembly of SNPs, indels and AS events') # ------------------------------------------------------------------------ # Define allowed options # ------------------------------------------------------------------------ parser.add_argument("-r", action="append", dest="readfiles", help="input fasta/q read files or compressed (.gz) fasta/q files (mutiple, such as \"-r file1 -r file2...\") ") parser.add_argument('-k', action="store", dest="kval", type=int, default=41, help="k-mer size (default=41)") parser.add_argument('-b', action="store", dest="bval", type=int, default=5, help="maximum number of branching nodes (default 5)") parser.add_argument('-l', action="store", dest="llmax", type=int, default=0, help="maximal length of the shorter path (default: 2k+1)") parser.add_argument('-m', action = "store", dest = "LL_MIN", default = 0, help = "minimum length of the shorter path (default 2k-8)") parser.add_argument('-M', action = "store", dest = "UL_MAX", default = 1000000, help = "maximum length of the longest path (default 1000000), skipped exons longer than UL_MAX are not reported") parser.add_argument('-g', action="store", dest="graph_prefix", default="", help="path and prefix to pre-built de Bruijn graph (suffixed by .edges/.nodes)\n \ if jointly used with -r, graph used to find bubbles and reads used for quantification") parser.add_argument('-o', action="store", dest="out_dir", default="results", help="path to store the results and the summary log file (default = ./results)") parser.add_argument('-d', action="store", dest="path_to_tmp", default=None, help="specific directory (absolute path) where to build temporary files (default temporary directory otherwise)") parser.add_argument('-t', action="store", dest="nbprocs", type=int, default=1, help="number of cores (must be <= number of physical cores)") parser.add_argument('-s', action="store", dest="output_snps", default = "0", help="0, 1 or 2. Changes which types of SNPs will be output. If 0 (default), will not output SNPs. If 1, will output Type0a-SNPs. If 2, will output Type0a and Type0b SNPs (warning: this option may increase a lot the running time. You might also want to try the experimental algorithm here)") parser.add_argument('-v', action="store_true", dest="verbose", help="Verbose mode") parser.add_argument('-u', action="store_true", dest="keep_ubccs", help="keep the nodes/edges file for unfinished bccs") parser.add_argument('-c', action = "store", type = int, dest = "min_cov", default = 2, help="an integer, k-mers present strictly less than this number of times in the dataset will be discarded (default 2)") parser.add_argument('-C', action = "store", type = float, dest = "min_relative_cov", default = 0.05, help="a percentage from [0,1), edges with relative coverage below this number are removed (default 0.05)") parser.add_argument('-e', action = "store", dest = "min_edit_dist", default = 3, help="edit distance threshold, if the two sequences (paths) of a bubble have edit distance smaller than this threshold, the bubble is classified as an inexact repeat (default 3)") parser.add_argument('-y', action = "store", dest = "max_cycles", default = 100000000, help="maximal number of bubbles enumeration in each bcc. If exceeded, no bubble is output for the bcc (default 100M)") parser.add_argument('--mismatches', action = "store", dest = "mism", default = 2, type = int, help="Maximal number of substitutions authorized between a read and a fragment (for quantification only), default 2. If you increase the mismatch and use --counts think of increasing min_overlap too.") parser.add_argument('--mismatchesSNP', action = "store", dest = "mismSNP", default = 0, type = int, help="Maximal number of substitutions authorized between a read and a fragment (for quantification only) for SNP, default 0.") parser.add_argument('--counts', action = "store", dest = "countsMethod", default = "2", help="0,1 or 2 . Changes how the counts will be reported. If 0 : total counts, if 1: counts on junctions, if 2 (default): all counts. see User guide for more information ") parser.add_argument('--min_overlap', action = "store", dest = "minOverlap", default = 5, type=int, help="Set how many nt must overlap a junction to be counted by --counts option. Default=5. see User guide for more information ") parser.add_argument('--timeout', action='store', dest="timeout", default=TIMEOUT, help="max amount of time (in seconds) spent for enumerating bubbles in each bcc. If exceeded, no bubble is output for the bcc (default "+str(TIMEOUT)+")") parser.add_argument('--version', action='version', version='%(prog)s @PROJECT_VERSION@') parser.add_argument('--output-context', action="store_true", dest="output_context", default = False, help="Will output the maximum non-ambiguous context of a bubble") parser.add_argument('--output-path', action="store_true", dest="output_path", default = False, help="Will output the id of the nodes composing the two paths of the bubbles.") parser.add_argument('--output-branch-count', action="store_true", dest="output_branch_count", default = False, help="Will output the number of branching nodes in each path.") parser.add_argument('--keep-bccs', action="store_true", dest="keep_all_bccs", default = False, help="Keep the node/edges files for all bccs.") parser.add_argument('--not-experimental', action="store_false", dest="experimental", default = True, help="Do not use a new experimental algorithm that searches for bubbles by listing all paths.") parser.add_argument('--max-memory', action="store", dest="max_memory", default="unlimited", help="If you use the experimental algorithm, you must provide the maximum size of the process's virtual memory (address space) in megabytes (default unlimited). WARNING: this option does not work on Mac operating systems.") parser.add_argument('--keep-counts', action="store_true", dest="keep_counts", default = False, help="Keep the .counts file after the sequencing-errors-removal step.") parser.add_argument('--get-mapping-info', action="store_true", dest="get_mapping_info", default = False, help="Creates a file with the KissReads mapping information of the reads on the bubbles.") parser.add_argument('--stranded', action="store_true", dest="stranded", default = False, help="Execute kissreads in stranded mode.") parser.add_argument('--strandedAbsoluteThreshold', action = "store", dest = "strandedAbsoluteThreshold", default = 3, type=int, help="Sets the minimum number of reads mapping to a path of a bubble in a read set is needed to call a strand.") parser.add_argument('--strandedRelativeThreshold', action = "store", dest = "strandedRelativeThreshold", default = 0.75, help="If a strand is called for a path of a bubble in a read set, but the proportion of reads calling this strand is less than this threshold, then the strand of the path is set to '?' (any strand - not enough evidence to call a strand).") parser.add_argument('--keep-redundancy', action="store_true", dest="keep_rd", default = False, help="Keep the Type_1 redundant cycles in the result file.") parser.add_argument('--keep-low-complexity', action="store_true", dest="keep_lc", default = False, help="Keep the low-complexity Type_1 cycles in the result file.") parser.add_argument('--lc-entropy-threshold', action = "store", dest = "lc_ent", default = ENTROPYMAX, type=int, help="Cycles with a Shannon entropy value for their upper path below this value will be removed (use --keep-low-complexity to keep them).") parser.add_argument('--get-redundance-info', action="store_true", dest="get_rd_info", default = False, help="Creates files with informations on compressed redundant cycles.") parser.add_argument('--get-low-complexity-info', action="store_true", dest="get_lc_info", default = False, help="Creates a file with informations on removed low-complexity cycles.") parser.add_argument('--type1-only', action="store_true", dest="t1o", default = False, help="Only quantify Type 1 bubbles (alternative splicing events, MAJOR SPEED UP with -b > 10 BUT all other bubbles will not appear in the result file).") # ------------------------------------------------------------------------ # Parse and interpret command line arguments # ------------------------------------------------------------------------ options = parser.parse_args() # ------------------------------------------------------------------------ # Create output dir # ------------------------------------------------------------------------ outdir = options.out_dir if not os.path.exists(outdir): os.mkdir(outdir) # ------------------------------------------------------------------------ # Create the log file # ------------------------------------------------------------------------ global logFile, logFileName logFileName = outdir+"/kissplice_log_summary_"+time.strftime("%Y-%m-%d")+"_"+time.strftime("%H-%M-%S")+"_"+str(randint(0, 1000000)) logFile = open(logFileName, 'w') # ------------------------------------------------------------------------ # Print version and command line # ------------------------------------------------------------------------ print("\nThis is KisSplice, version @PROJECT_VERSION@\n") printlg("This is KisSplice, version @PROJECT_VERSION@\n") print("The command line was: " + ' '.join(sys.argv)) printlg("The command line was: " + ' '.join(sys.argv)) # ------------------------------------------------------------------------ # Parse input options # ------------------------------------------------------------------------ # check if the given read files indeed exist check_read_files(options.readfiles) readfiles = None only_graph = False if options.readfiles: if options.graph_prefix: # GRAPH + READS print("-r and -g options used together: ") printlg("-r and -g options used together: ") print("the graph will be used to find events, while reads files are used for checking read-coherency and coverage") printlg("the graph will be used to find events, while reads files are used for checking read-coherency and coverage") readfiles = ' '.join(map(str, options.readfiles)) else: if not options.graph_prefix: parser.print_usage() dieToFatalError("kissplice requires at least a read file or a pre-built graph") else: # GRAPH only_graph = True nobuild = False if options.graph_prefix: nobuild = True # --------------------------------------------------------- Output options output_snps = (int)(options.output_snps) if output_snps<0 or output_snps>2: print("-s is not 0, 1 or 2. Defaulting to 0.") printlg("-s is not 0, 1 or 2. Defaulting to 0.") output_snps = 0 print("Using the read files: ", readfiles) printlg("Using the read files: ", readfiles) print("Results will be stored in: ", os.path.abspath(options.out_dir)) printlg("Results will be stored in: ", os.path.abspath(options.out_dir)) print("Summary log file will be saved in: ", os.path.abspath(logFileName)) printlg("Summary log file will be saved in: ", os.path.abspath(logFileName)) print("\n") printlg("\n") # ------------------------------------------------------------- k-mer size kval = options.kval if kval%2 == 0: dieToFatalError("please use only odd value for k") #otherwise, DBG use k-1 and output context do not work # ------------------------------------- Maximal length of the shorter path if options.llmax != 0: LL_MAX = options.llmax else: LL_MAX = 2 * kval + 1 # The following are not optional but work along with llmax UL_MAX = options.UL_MAX # Defines maximum upper and lower path bounds if options.LL_MIN != 0: LL_MIN= options.LL_MIN else: LL_MIN = 2 * kval - 8 min_ll_max = 2 * kval + 1 if LL_MAX < min_ll_max: dieToFatalError("maximal length of the shorter path (" + str(LL_MAX) + ") should be >= 2k+1 =" + str(min_ll_max) + ")") #-------------------------------- fix LL_MIN, LL_MAX and UL_MAX -------------------------------------- LL_MIN = int(LL_MIN)-2 LL_MAX = int(LL_MAX)-2 UL_MAX = int(UL_MAX)-2 # ------------------------------------------------------- Other parameters min_cov = options.min_cov min_edit_dist = options.min_edit_dist max_cycles = options.max_cycles countsMethod = options.countsMethod minOverlap = options.minOverlap # ======================================================================== # Construct intermediate and output file names # ======================================================================== workdir = mkdirTmp(options.path_to_tmp) infix_name = "" # will be the central part of the output file names if options.graph_prefix: graphfile = options.graph_prefix if options.readfiles: for file in options.readfiles: justfilename = file.split("/")[-1].split(".")[0] #remove what is before the "/" and what is after the "." infix_name += justfilename+"_" infix_name = infix_name[0:200] # Truncate it to contain at most 200 characteres infix_name += "k" + str(kval) if not options.graph_prefix: graphfile = options.out_dir+"/graph_"+infix_name # Output graph file else: infix_name = graphfile.split("/")[-1].split(".")[0] #remove what is before the "/" and what is after the "." # ======================================================================== # RUN # ======================================================================== # ------------------------------------------------------------------------ # Build De Bruijn Graph # ------------------------------------------------------------------------ if not nobuild: t = time.time() build_graph(internal_bindir, workdir, readfiles, kval, graphfile, min_cov, options.nbprocs, options.verbose) print("Elapsed time: ", round(time.time() - t,1), " seconds") printlg("Elapsed time: ", round(time.time() - t,1), " seconds") # ------------------------------------------------------------------------ # Error removal # ------------------------------------------------------------------------ t = time.time() if float(options.min_relative_cov) > 0.0001: exec_error_removal = error_removal(internal_bindir, graphfile, nobuild, options.min_relative_cov, options.verbose) else: exec_error_removal = False print("Elapsed time: ", round(time.time() - t,1), " seconds") printlg("Elapsed time: ", round(time.time() - t,1), " seconds") # ------------------------------------------------------------------------ # Decompose and simplify DBG # ------------------------------------------------------------------------ t = time.time() run_modules(internal_bindir, workdir, graphfile, kval, options.min_relative_cov, options.verbose, options.output_context, exec_error_removal) print("Elapsed time: ", round(time.time() - t,1), " seconds") printlg("Elapsed time: ", round(time.time() - t,1), " seconds") # ------------------------------------------------------------------------ # Enumerate Bubbles # ------------------------------------------------------------------------ t = time.time() enumerate_all_bubbles(internal_bindir, workdir, outdir, kval, options.bval, output_snps, min_edit_dist, max_cycles, \ UL_MAX, LL_MAX, LL_MIN, float(options.timeout), options.nbprocs, options.verbose, \ options.output_context, options.output_path, options.output_branch_count, options.experimental, options.max_memory) print("Elapsed time: ", round(time.time() - t,1), " seconds") printlg("Elapsed time: ", round(time.time() - t,1), " seconds") # ------------------------------------------------------------------------ # Sort and remove redundancy/low-complexity bubbles (optionnal), only keep type_1 events (optionnal) # ------------------------------------------------------------------------ t = time.time() nb = sort_all_bubbles(internal_bindir, readfiles, workdir, outdir, kval, output_snps, infix_name, not only_graph, options.verbose) filesToMove=redundancyAndLowComplexityRemoval(workdir, "all_bcc_type1234_"+str(kval), options.keep_rd, options.keep_lc, options.lc_ent, options.get_rd_info, options.get_lc_info, options.t1o, kval) for fToMove in filesToMove: shutil.move("/".join([workdir,fToMove]), "/".join([outdir,fToMove])) print("Elapsed time: "+str(round(time.time() - t,1))+" seconds") printlg("Elapsed time: "+str(round(time.time() - t,1))+" seconds") # ------------------------------------------------------------------------ # Check read coverage (optionnal) # ------------------------------------------------------------------------ t = time.time() if not only_graph: nb = check_read_coverage_and_sort_all_bubbles(internal_bindir, readfiles, workdir, outdir, kval, output_snps, infix_name, countsMethod, minOverlap, options.mism, options.mismSNP, options.get_mapping_info, options.stranded, options.strandedAbsoluteThreshold, options.strandedRelativeThreshold, options.nbprocs, options.verbose) print("Elapsed time: ", round(time.time() - t,1), " seconds\n") printlg("Elapsed time: ", round(time.time() - t,1), " seconds\n") if only_graph: print("\n\n \t\t ******** We are done, final results are in files "+outdir+"/results_"+infix_name+"_type_*.fa **********") printlg("\n\n \t\t ******** We are done, final results are in files "+outdir+"/results_"+infix_name+"_type_*.fa **********") else: print("\n\n \t\t ******** We are done, final coherent results are in files "+outdir+"/results_"+infix_name+"_coherents_type_*.fa ********** ") printlg("\n\n \t\t ******** We are done, final coherent results are in files "+outdir+"/results_"+infix_name+"_coherents_type_*.fa ********** ") print(" \t\t ******** All non read coherent results are in files "+outdir+"/results_"+infix_name+"_uncoherent.fa ****** \n\n") printlg(" \t\t ******** All non read coherent results are in files "+outdir+"/results_"+infix_name+"_uncoherent.fa ****** \n\n") # ------------------------------------------------------------------------ # Manage BCCs # ------------------------------------------------------------------------ if len(unfinished_bccs) != 0: print("\t\t ******** There were " + str(len(unfinished_bccs)) + " BCCs with unfinished enumeration ********") printlg("\t\t ******** There were " + str(len(unfinished_bccs)) + " BCCs with unfinished enumeration ********") if not options.keep_ubccs and not options.keep_all_bccs: print("\t\t ******** Re-run with `-u` to retrieve the unfinished components ********\n") printlg("\t\t ******** Re-run with `-u` to retrieve the unfinished components ********\n") if options.keep_ubccs: bcc_dir = "/unfinished_bcc" print("\t\t Backup files for the unfinished BCCs are in " + outdir + bcc_dir + "\n") printlg("\t\t Backup files for the unfinished BCCs are in " + outdir + bcc_dir + "\n") save_bccs_from_list(unfinished_bccs, bcc_dir, internal_bindir, workdir, outdir, options.verbose) if options.keep_all_bccs: bcc_dir = "/bcc" print("\t\t Edge and node files of all BCCs are in " + outdir + bcc_dir + "\n") printlg("\t\t Edge and node files of all BCCs are in " + outdir + bcc_dir + "\n") all_bccs = find_bcc_ids_ordered_by_size(workdir)[1] save_bccs_from_list(all_bccs, bcc_dir, internal_bindir, workdir, outdir, options.verbose) if options.output_path: # move paths file to outdir shutil.move(workdir+"/all_paths_k"+str(kval), outdir + "/all_paths_k"+str(kval)) # ------------------------------------------------------------------------ # Output number of events of each type # ------------------------------------------------------------------------ print("\t\t TYPES:") printlg("\t\t TYPES:") if output_snps!=0: print("\t\t\t 0a: Single SNPs, Inexact Repeats or sequencing substitution errors, "+str(int(nb[0]))+" found") printlg("\t\t\t 0a: Single SNPs, Inexact Repeats or sequencing substitution errors, "+str(int(nb[0]))+" found") if output_snps==2: print("\t\t\t 0b: Multiple SNPs, Inexact Repeats or sequencing substitution errors, "+str(int(nb[1]))+" found") printlg("\t\t\t 0b: Multiple SNPs, Inexact Repeats or sequencing substitution errors, "+str(int(nb[1]))+" found") else: print("\t\t\t 0b: Run with -s 2 to also search for Multiple SNPs (warning: this option may increase a lot the running time)") printlg("\t\t\t 0b: Run with -s 2 to also search for Multiple SNPs (warning: this option may increase a lot the running time)") else: print("\t\t\t 0: Run with -s option set to 1 or 2 to also search for SNPs") printlg("\t\t\t 0: Run with -s option set to 1 or 2 to also search for SNPs") print("\t\t\t 1: Alternative Splicing Events, "+str(int(nb[2]))+" found") printlg("\t\t\t 1: Alternative Splicing Events, "+str(int(nb[2]))+" found") print("\t\t\t 2: Inexact Tandem Repeats, "+str(int(nb[3]))+" found") printlg("\t\t\t 2: Inexact Tandem Repeats, "+str(int(nb[3]))+" found") print("\t\t\t 3: Short Indels (<3nt), "+str(int(nb[4]))+" found") printlg("\t\t\t 3: Short Indels (<3nt), "+str(int(nb[4]))+" found") print("\t\t\t 4: All others, composed by a shorter path of length > 2k not being a SNP, "+str(int(nb[5]))+" found") printlg("\t\t\t 4: All others, composed by a shorter path of length > 2k not being a SNP, "+str(int(nb[5]))+" found") print("\n\n \t\t ******** A summary of the execution can be found in the log file: " + os.path.abspath(logFileName) + "**********") printlg("\n\n \t\t ******** A summary of the execution can be found in the log file: " + os.path.abspath(logFileName) + "**********") # ------------------------------------------------------------------------ # Clean tmp directory # ------------------------------------------------------------------------ logFile.close() cleanTmp(workdir) if __name__ == '__main__': main() kissplice-2.6.2/man/000077500000000000000000000000001423273700400142645ustar00rootroot00000000000000kissplice-2.6.2/man/CMakeLists.txt000066400000000000000000000005461423273700400170310ustar00rootroot00000000000000# Fill manpage template with version and date # TODO maybe use tools like help2man to automate manpage, considering this is mostly a --help message ? configure_file("${CMAKE_CURRENT_SOURCE_DIR}/kissplice.in.1" "${CMAKE_CURRENT_BINARY_DIR}/kissplice.1" @ONLY) install(FILES "${CMAKE_CURRENT_BINARY_DIR}/kissplice.1" DESTINATION "${CMAKE_INSTALL_MANDIR}/man1")kissplice-2.6.2/man/kissplice.in.1000066400000000000000000000117351423273700400167500ustar00rootroot00000000000000.TH KISSPLICE "1" "@CONFIGURE_DATE@" "kissplice @PROJECT_VERSION@" "User Commands" .SH NAME kisSplice \- local assembly of SNPs, indels and AS events .SH SYNOPSIS kissplice \-h .br kissplice [OPTION] [\-r READFILES] .SH DESCRIPTION Detects alternative splicing events and other kinds of polymorphisms from READFILES (in FASTA or FASTQ format). .SH OPTIONS .TP .B \-h, \-\-help Show this help message and exit. .TP \fB\-r\fR READFILES Input fasta/q read files or compressed (.gz) fasta/q files (mutiple, such as "-r file1 -r file2...") .TP \fB\-k\fR KVAL k\-mer size (default=41). .TP \fB\-b\fR BVAL Maximum number of branching nodes (default: 5) .TP \fB\-l\fR LLMAX Maximal length of the shorter path (default: 2k\+1). .TP \fB\-m\fR LL_MIN Minimum length of the shorter path (default 2k\-8). .TP \fB\-M\fR UL_MAX Maximum length of the longest path (default: 1000000), skipped exons longer than UL_MAX are not reported. .TP \fB\-g\fR GRAPH_PREFIX Path and prefix to pre\-built de Bruijn graph (suffixed by .edges/.nodes) if jointly used with \fB\-r\fR, graph used to find bubbles and reads used for quantification. .TP \fB\-o\fR OUT_DIR Path to store the results (default = ./results). .TP \fB\-d\fR PATH_TO_TMP Specific directory (absolute path) where to build temporary files (default temporary directory otherwise). .TP \fB\-t\fR NBPROCS Number of cores (must be <= number of physical cores). .TP \fB\-s\fR OUTPUT_SNPS 0, 1 or 2. Changes which types of SNPs will be output. If 0 (default), will not output SNPs. If 1, will output Type0a-SNPs. If 2, will output Type0a and Type0b SNPs (warning: this option may increase a lot the running time.). .TP \fB\-v\fR Verbose mode. .TP \fB\-u\fR Keep the nodes/edges file for unfinished bccs. .TP \fB\-c\fR MIN_COV Discard k\-mers tha are present strictly less than this number of times in the dataset. (default 2). .TP \fB\-C\fR MIN_RELATIVE_COV Discard edges with relative coverage below MIN_RELATIVE_COV expressed as a percentage in [0,1). (default 0.05). .TP \fB\-e\fR MIN_EDIT_DIST Classify as inexact repeats those bubbles whose paths' edit distance is smaller than MIN_EDIT_DIST (default 3). .TP \fB\-y\fR MAX_CYCLES Maximal number of bubble enumerations in each bcc. If exceeded, no bubble is output for the bcc (default: 100000000). .TP \fB\-\-mismatches\fR NB_MISMATCHES Maximal number of substitutions authorized between a read and a fragment (for quantification only), default 2. If you increase the mismatch and use \fB\-\-counts\fR think of increasing min_overlap too. .TP \fB\-\-counts\fR COUNTS_TYPE Changes how the counts will be reported. If 0: total counts, if 1: counts on junctions, if 2 (default): all counts. .TP \fB\-\-min_overlap\fR MIN_OVERLAP Sets how many nt must overlap a junction to be counted by --counts option (default: 5). .TP \fB\-\-timeout\fR TIMEOUT Max amount of time (in seconds) spent for enumerating bubbles in each bcc. If exceeded, no bubble is output for the bcc (default 100000). .TP \fB\-\-version\fR Display program's version number and exit. .TP \fB\-\-output-context\fR Will output the maximum non-ambiguous context of a bubble. .TP \fB\-\-output-path\fR Will output the id of the nodes composing the two paths of the bubbles. .TP \fB\-\-output-branch-count\fR Will output the number of branching nodes in each path. .TP \fB\-\-keep-bccs\fR Keep the node/edges files for all bccs. .TP \fB\-\-not-experimental\fR Do not use a new experimental algorithm that searches for bubbles by listing all paths. .TP \fB\-\-max-memory MAX_MEMORY\fR If you use the experimental algorithm, you must provide the maximum size of the process's virtual memory (address space) in megabytes (default unlimited). .TP \fB\-\-keep-counts\fR Keep the .counts file after the sequencing-errors-removal step. .TP \fB\-\-get-mapping-info\fR Creates a file with the KissReads mapping information of the reads on the bubbles. .TP \fB\-\-stranded\fR Execute kissreads in stranded mode. .TP \fB\-\-strandedAbsoluteThreshold\fR Sets the minimum number of reads mapping to a path of a bubble in a read set is needed to call a strand. .TP \fB\-\-strandedRelativeThreshold\fR If a strand is called for a path of a bubble in a read set, but the proportion of reads calling this strand is less than this threshold, then the strand of the path is set to '?' (any strand - not enough evidence to call a strand). .TP \fB\-\-keep-redundancy\fR Keep the Type_1 redundant cycles in the result file. .TP \fB\-\-keep-low-complexity\fR Keep the low-complexity Type_1 cycles in the result file. .TP \fB\-\-lc-entropy-threshold\fR Cycles with a Shannon entropy value for their upper path below this value will be removed (use --keep-low-complexity to keep them). .TP \fB\-\-get-redundance-info\fR Creates files with informations on compressed redundant cycles. .TP \fB\-\-get-low-complexity-info\fR Creates a file with informations on removed low-complexity cycles. .TP \fB\-\-type1-only\fR Only quantify Type 1 bubbles (alternative splicing events, MAJOR SPEED UP with -b > 10 BUT all other bubbles will not appear in the result file).kissplice-2.6.2/modules/000077500000000000000000000000001423273700400151615ustar00rootroot00000000000000kissplice-2.6.2/modules/BubbleEnumeration.cpp000066400000000000000000000703161423273700400212760ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #include #include #include #include #include #include #include #include #include #include "LabelledCEdge.h" #include "WeightedDigraph.h" #include "CGraph.h" #include "BubbleEnumeration.h" #include "Utils.h" #include #define MAX_DIST 10000000 #define MAX 1024 using namespace std; int nbBubbles = 0; //these 3 variables represent beta, a1 and a2. We have a1Global and a2Global so that it does not confuse with some locals a1 and a2 int beta = 0; int a1Global = 0; int a2Global = 0; int MAX_BUBBLES; int UL_MAX; int LL_MAX; int LL_MIN; int MIN_DIST; int BUBBLE_COUNT_OFFSET; int MAX_BRANCHES; int MAX_MEMORY; int OUTPUT_SNPS; bool OUTPUT_CONTEXT; bool OUTPUT_PATH; bool OUTPUT_BRANCH; bool EXPERIMENTAL_ALG; // OUTPUT FILE, One for SNPs, One for other type of Bubble FILE *seq_output_file_type0; FILE *seq_output_file_type1234; FILE *path_output_file; string comment = ""; vector seqs; int k_value; int nb_nodes; //////////////////////////////////////////////////////////////////////////////////////////////////////////////// /* Returns the sequence for node "i", stored in the seqs vector. There is a catch, the vector does not store the sequence of the complementary node. The function has to convert it. */ string getSeq(int i) { return (i < (int)seqs.size()) ? string(seqs[i]) : reverse_complement(string(seqs[i % (int)seqs.size()])); } /*! * \brief Return the path sequence ( with or without context) * \param path: nodes ids path * \param all_nodes : output_context boolean * \param output: if seq gotten for output, then there is one nt more on each side of the path * to detect various switching nodes */ string path2seq(vector& path, bool all_nodes, bool output) { string pseq = getSeq( path[0] ); for (int i = 1; i < (int)path.size(); i++) { pseq += getSeq(path[i]).substr(k_value-1); } if (!all_nodes) { int len_first; int len_last ; if ( output ) { len_first = getSeq( path[0] ).size() - (k_value ); len_last = getSeq( path[(int)path.size()-1] ).size() - (k_value ); } else { len_first = getSeq( path[0] ).size() - (k_value ); len_last = getSeq( path[(int)path.size()-1] ).size() - (k_value ); } return pseq.substr(len_first, (int) pseq.size() - (len_first + len_last) ); } return pseq; } void print_path(FILE* stream, vector& path) { for (int i = 0; i < (int)path.size(); i++) { fprintf(stream, " %d", path[i]); } fprintf(stream, "\n"); } void print_formated_path(FILE* stream, string& bcc, int cycle_num, string& type, string path_name, int len, string& bcount, string& seq) { fprintf(stream, ">%s|Cycle_%d|Type_%s|%s_length_%d%s\n", bcc.c_str(), cycle_num, type.c_str(), path_name.c_str(), len, bcount.c_str()); fprintf(stream, "%s\n", seq.c_str()); } void output_bubble(FILE* stream, string& upper, int upper_b, string& lower, int lower_b, string type, int num, int contextFirst, int contextLast) { // passing in lower case context if(contextFirst != 0 || contextLast != 0) { upper = toLowerContext( upper, contextFirst, contextLast); lower = toLowerContext( lower, contextFirst, contextLast); } string upper_bcount = "", lower_bcount = ""; if (OUTPUT_BRANCH) { upper_bcount = "_branches_" + to_string(upper_b); lower_bcount = "_branches_" + to_string(lower_b); } print_formated_path(stream, comment, num, type, "upper_path", (int)upper.size() - (contextFirst + contextLast), upper_bcount, upper); print_formated_path(stream, comment, num, type, "lower_path", (int)lower.size() - (contextFirst + contextLast), lower_bcount, lower); } /* * for output node-ids option */ void output_bubble(FILE* stream, vector& upper, int upper_b, vector& lower, int lower_b, string type, int num) { fprintf(stream, ">%s|Cycle_%d|Type_%s|upper_path_length_%d\n", comment.c_str(), num, type.c_str(), (int)upper.size()); print_path(stream, upper); fprintf(stream, ">%s|Cycle_%d|Type_%s|lower_path_length_%d\n", comment.c_str(), num, type.c_str(), (int)lower.size()); print_path(stream, lower); } string classify_bubble(vector& p1_nodes, vector& p2_nodes) { string upper_seq = path2seq(p1_nodes, false, false); string lower_seq = path2seq(p2_nodes, false, false); /* truncated -1 nt left and -1nt right, they only serve to distinguish Swithcing Nodes, it has nothing to do with quantification */ upper_seq = upper_seq.substr(1, upper_seq.size() - 2); lower_seq = lower_seq.substr(1, lower_seq.size() - 2); if (upper_seq.size() < lower_seq.size()) swap(upper_seq, lower_seq); string type; int u_len = (int) upper_seq.size(), l_len = (int) lower_seq.size(); if (l_len <= (2 * k_value - 2)) // lower path of at most 2k-2: splicing, repeats, and indels { int d1, d2; if ((u_len - l_len) > 0 && (((u_len - l_len) <= 2) || ((u_len - l_len) == 4) || ((u_len - l_len) == 5))) //indel type = "3"; else if ( (d1 = edit_distance(upper_seq.c_str(), l_len, lower_seq.c_str(), l_len, sizeof (char), comp)) <= MIN_DIST || (d2 = edit_distance(upper_seq.c_str() + u_len - l_len, l_len, lower_seq.c_str(), l_len, sizeof (char), comp)) <= MIN_DIST) // tandem repeat type = "2"; else //splicing type = "1"; } else if (u_len == l_len && // SNP //specific condition for Type_0a ((u_len == 2 * k_value - 1) || //specific condition for Type_0b, at most 10% differences in the variable region. Arbitrary TODO: check this (hamming_distance(upper_seq.c_str()+(k_value-1), l_len-(2*k_value)+2, lower_seq.c_str()+(k_value-1), l_len-(2*k_value)+2, sizeof (char), comp) <= (0.1 * (l_len-(2*k_value)+2))))) { if (u_len > 2 * k_value - 1) // multiple SNPs type = "0b"; else type = "0a"; } else if (l_len <= LL_MAX) // others with lower path of length bigger than 2k-2 and smaller or equal to LL_MAX type = "4"; else //here we have bubbles such that the lower path is bigger than LL_MAX and is not a Type0b. This can only happen by default if we were searching for Type0bs and a bubble is not a Type0b type = "undefined"; return type; } /////////////////////////////////////////////////////////////////////////////////////////////////////////// set in_sub; bool contains(set &s, int elem) { return s.find(elem) != s.end(); } /* Used by the heap, as the pair . */ typedef pair ii; /* Dijsktra's algorithm. Returns the distance (shortest path) from source to every node. The results are in dist. */ void dijkstra(int max_dist, int source, map& dist, WeightedDigraph& G) { priority_queue, greater > Q; dist[source] = 0; Q.push(ii(0, source)); while (!Q.empty()) { ii top = Q.top(); Q.pop(); int v = top.second; for (int i = 0; i < (int)G.adj_list[v].size(); i++) if (!G.adj_list[v][i].removed) { int w = G.adj_list[v][i].node, cost = G.adj_list[v][i].cost; if (!G.removed[w] && contains(in_sub, w)) { if (dist.find(w) == dist.end()) dist[w] = MAX_DIST; if (dist[w] > dist[v] + cost) { dist[w] = dist[v] + cost; Q.push(ii(dist[w], w)); } } } } } /* Checks if the distance from s to t is smaller than max_dist */ bool dist_st(int max_dist, int s, int t, WeightedDigraph& G) { map dist; dist[t] = MAX_DIST; dijkstra(max_dist, s, dist, G); return dist[t] <= max_dist + G.node_cost[t]; } /* Check if there exist a pair compatible paths from s1, s2 respecting a1, a2. Check the paper for a precise definition of pair of compatible paths. */ bool testPairCompatible(int s1, int a1, int s2, int a2, WeightedDigraph& G) { map dist1, dist2; dijkstra(a1, s1, dist1, G); dijkstra(a2, s2, dist2, G); map::iterator it; for (it = dist1.begin(); it != dist1.end(); it++) if (dist2.find(it->first) != dist2.end()) { int i = it->first; if (dist1[i] <= (a1 + G.node_cost[i]) && dist2[i] <= (a2 + G.node_cost[i])) return true; } return false; } /* In-degree is equal to out-degree of the completary node, because it's a de Bruijn graph.*/ int inDegree(int node, WeightedDigraph& G) { if (node < nb_nodes) return G.outDegree(node + nb_nodes); else return G.outDegree(node - nb_nodes); } int nbBranchingNodes(vector& p_nodes, WeightedDigraph& G) { int nb = 0; // Disregard the initial and final nodes. for (int i = 1; i < (int)p_nodes.size()-1; i++) if (G.outDegree(p_nodes[i]) != 1 || inDegree(p_nodes[i],G) != 1) nb++; return nb; } void processBubbleFound(vector &p1_nodes, vector &p2_nodes, WeightedDigraph &G) { string type = classify_bubble(p1_nodes, p2_nodes); if (type != "undefined" && (type[0] != '0' || OUTPUT_SNPS)) { string upper_seq = path2seq(p1_nodes, OUTPUT_CONTEXT, true); string lower_seq = path2seq(p2_nodes, OUTPUT_CONTEXT, true); int upper_b = nbBranchingNodes(p1_nodes, G), lower_b = nbBranchingNodes(p2_nodes, G); bool swp = false; if (upper_seq.size() < lower_seq.size()) { swap(upper_seq, lower_seq); swap(upper_b, lower_b); swp = true; } FILE *selection = (type[0] == '0') ? seq_output_file_type0 : seq_output_file_type1234; // Writting in different files if type 0 or else if (OUTPUT_CONTEXT) { // getting the context: size of the first (or last ) node - (k-1) right int contextLeft = getSeq(p1_nodes[0]).size() - (k_value); int contextRight = getSeq(p1_nodes[(int) p1_nodes.size() - 1]).size() - (k_value); output_bubble(selection, upper_seq, upper_b, lower_seq, lower_b, type, BUBBLE_COUNT_OFFSET + nbBubbles, contextLeft, contextRight); } else { output_bubble(selection, upper_seq, upper_b, lower_seq, lower_b, type, BUBBLE_COUNT_OFFSET + nbBubbles, 0, 0); // size of the context if null } if (OUTPUT_PATH) { output_bubble(path_output_file, swp ? p2_nodes : p1_nodes, upper_b, swp ? p1_nodes : p2_nodes, lower_b, type, BUBBLE_COUNT_OFFSET + nbBubbles); } } nbBubbles++; if (nbBubbles > MAX_BUBBLES) exit(15); } /*! * \brief list all pair of compatible paths (see paper), classify the events found * and output them * \param s1 * \param a1 * \param p1 * \param p1_nodes * \param s2 * \param a2 * \param p2 * \param p2_nodes * \param G * * * List all pair of compatible paths from s1,s2 respecting a1,a2. It's a recursive algorithm based on the bipartition method. Check the paper for a full description. */ void listPairCompatible(int s1, int a1, int p1, vector& p1_nodes, int s2, int a2, int p2, vector& p2_nodes, WeightedDigraph& G) { if (nbBranchingNodes(p1_nodes, G) > MAX_BRANCHES || nbBranchingNodes(p2_nodes, G) > MAX_BRANCHES) return; if (s1 == s2 && (p1_nodes.size() != 1 || p2_nodes.size() != 1)) { if ( p1 >= (beta + G.node_cost[s1]) && p2 >= (beta + G.node_cost[s2]) ) { processBubbleFound(p1_nodes, p2_nodes, G); } return; } if (G.adjListSz(s1) == 0 && G.adjListSz(s2) == 0) return; int u = (G.adjListSz(s1) != 0) ? s1 : s2; G.removed[u] = true; for (int i = 0; i < (int)G.adj_list[u].size(); i++) { int v = G.adj_list[u][i].node, cost = G.adj_list[u][i].cost; if (!G.removed[v] && !G.adj_list[u][i].removed && contains(in_sub, v)) { if (u == s1 && testPairCompatible(v, a1 - cost, s2, a2, G)) { p1_nodes.push_back(v); listPairCompatible(v, a1 - cost, p1 + cost, p1_nodes, s2, a2, p2, p2_nodes, G); p1_nodes.pop_back(); } else if (u == s2 && testPairCompatible(s1, a1, v, a2 - cost, G)) { p2_nodes.push_back(v); listPairCompatible(s1, a1, p1, p1_nodes, v, a2 - cost, p2 + cost, p2_nodes, G); p2_nodes.pop_back(); } } } G.removed[u] = false; if ((u == s1 && p1 >= (beta + G.node_cost[s1])) || (u == s2 && p2 >= (beta + G.node_cost[s2]))) { vector adj_u; adj_u.swap(G.adj_list[u]); if ((u == s1 && dist_st(a2, s2, s1, G)) || (u == s2 && dist_st(a1, s1, s2, G))) listPairCompatible(s1, a1, p1, p1_nodes, s2, a2, p2, p2_nodes, G); G.adj_list[u].swap(adj_u); } } void bfs(WeightedDigraph &G, int source, set &in_sub) { queue Q; map dist; Q.push(source); dist[source] = 0; while (!Q.empty()) { int u = Q.front(); Q.pop(); in_sub.insert(u); int d = dist[u]; for (int i = 0; i < (int)G.adj_list[u].size(); i++) { int v = G.adj_list[u][i].node; if (!contains(in_sub, v)) { Q.push(v); dist[v] = d + 1; } } // We don't compress non-branching paths to an edge, but to a // vertex, this means that each non-branching vertex maybe // followed by a branching vertex. Implying that a path with // MAX_BRANCHES contains at most 2*MAX_BRANCHES+1 // vertices. However, we don't count the first and last vertices // (that are always branching), so the length is 2 * // (MAX_BRANCHES+1). if (d > 2*(MAX_BRANCHES+1)) return; } dist.clear(); } /* List all bubbles satisfying the path constraints. It does so using listPairCompatible function. */ void listAllBubbles(WeightedDigraph& G, int k_value, int UL_MAX, int LL_MAX, int LL_MIN) { //local a1 and a2 int a1 = LL_MAX - (k_value - 1), a2 = UL_MAX - (k_value - 1); beta = LL_MIN - (k_value - 1); for (int v = 0; v < (int)G.adj_list.size(); v++) { in_sub.clear(); bfs(G, v, in_sub); vector p1_nodes, p2_nodes; p1_nodes.push_back(v); p2_nodes.push_back(v); //checks if the user wants to output type0b or not if (OUTPUT_SNPS!=2) { //no, call normally listPairCompatible(v, a1, 0, p1_nodes, v, a2, 0, p2_nodes, G); }else { //yes. With this algorithm, maybe the only way to do this is to reexecute it passing a2 as LL_MIN and filtering the bubbles found s.t. lower path length > a1 listPairCompatible(v, a2, 0, p1_nodes, v, a2, 0, p2_nodes, G); } } } ///////////////////////////////////////////////////////////////////////////////////////////////////// void printSummary( FILE* stream, int num_bubbles ) { fprintf( stream, "============================================================================\n" ); fprintf( stream, "Summary of results\n" ); fprintf( stream, "============================================================================\n" ); fprintf( stream, "No of bubbles: %d\n", num_bubbles ); fprintf( stream, "============================================================================\n" ); } void printUsageAndExit( char * name ) { fprintf( stderr, "Usage: %s infofile contents_file_edges contents_file_nodes basename_edges basename_nodes number_to_read k_value output_prefix edit_distance_threshold comment numbering_offset [-u UL_MAX] [-L LL_MAX] [-l LL_MIN] [-M MAX_BUBBLES] [-s] [-p] [-c] [-b] [-e MAX_MEMORY]\n", name ); fprintf( stderr, "\t [-u UL_MAX] Maximal length of the upper path of each bubble. Default: 1000000\n" ); fprintf( stderr, "\t [-L LL_MAX] Maximal length of the lower path of each bubble. Default: 2k-1\n" ); fprintf( stderr, "\t [-l LL_MIN] Minimal length of the lower path of each bubble. Default: 2k-8\n" ); fprintf( stderr, "\t [-M MAX_BUBBLES] Stop the process after this number of bubbles. Default: 10000\n" ); fprintf( stderr, "\t [-b MAX_BRANCHES] Maximum number of branches for each bubble. Default: 5\n"); fprintf( stderr, "\t [-v] Outputs the number of branching nodes in each path\n"); fprintf( stderr, "\t [-e MAX_MEMORY] Use an experimental algorithm that find bubbles by listing paths. You must provide the maximum size of the process's virtual memory (address space) in megabytes.\n"); exit( EXIT_FAILURE ); } //Path-enumeration algorithm to list bubbles - forward declaration void listAllBubblesUsingPath(WeightedDigraph& G); int main( int argc, char** argv ) { if ( argc < 12 ) printUsageAndExit( argv[0] ); vector label; vector allEdges; string output_prefix = argv[8]; k_value = atoi(argv[7]); seq_output_file_type0 = fopen( string(output_prefix + "_type0.fa").c_str(), "w" ); seq_output_file_type1234 = fopen( string(output_prefix + "_type1234.fa").c_str(), "w" ); MIN_DIST = atoi(argv[9]); comment = argv[10]; BUBBLE_COUNT_OFFSET = atoi(argv[11]); LL_MAX = 2 * k_value - 1; LL_MIN = 2 * k_value - 10; UL_MAX = 1000000; MAX_BUBBLES = 10000; MAX_BRANCHES = 5; OUTPUT_CONTEXT = false; OUTPUT_SNPS = 0; OUTPUT_PATH = false; OUTPUT_BRANCH = false; EXPERIMENTAL_ALG = false; MAX_MEMORY = 0; int required_sequence = atoi( argv[6] ); int temoin; while ( (temoin = getopt ( argc-11, &argv[11], "u:L:l:M:b:e:s:cpv" )) != -1 ) { switch ( temoin ) { case 'u' : { UL_MAX = atoi( optarg ); break; } case 'L' : { LL_MAX = atoi( optarg ); break; } case 'l' : { LL_MIN = atoi( optarg ); break; } case 'M': { MAX_BUBBLES = atoi( optarg ); break; } case 'b': { MAX_BRANCHES = atoi( optarg ); break; } case 's': { OUTPUT_SNPS = atoi( optarg ); switch (OUTPUT_SNPS) { case 0: fprintf(stderr, "Will not output SNPs and sequencing errors\n" ); break; case 1: fprintf(stderr, "Will output Type0a-SNPs\n" ); break; case 2: fprintf(stderr, "Will output Type0a and Type0b SNPs\n" ); break; } break; } case 'c': { OUTPUT_CONTEXT = true; fprintf(stderr, "Will output bubble contexts!\n"); break; } case 'p': { OUTPUT_PATH = true; path_output_file = fopen( string(output_prefix + ".path").c_str(), "w" ); break; } case 'v': { OUTPUT_BRANCH = true; break; } case 'e': { EXPERIMENTAL_ALG = true; if (strcmp(optarg, "unlimited") == 0) MAX_MEMORY = -1; else MAX_MEMORY = atoi( optarg ); break; } default: { printUsageAndExit( argv[0] ); } } } fprintf(stdout, "\t Enumerating bubbles with at most %d branching nodes in each path!\n", MAX_BRANCHES); if (EXPERIMENTAL_ALG) { fprintf(stdout, "\t Using the experimental algorithm with maximum memory = "); if (MAX_MEMORY==-1) printf("unlimited.\n"); else printf("%d MB.\n", MAX_MEMORY); struct rlimit vmLimit; vmLimit.rlim_cur = vmLimit.rlim_max = (MAX_MEMORY==-1 ? RLIM_INFINITY : MAX_MEMORY * 1024 * 1024); setrlimit(RLIMIT_AS, &vmLimit); } EdgeLoader edgeloader(allEdges); NodeLoader nodeloader(seqs, label); bool atleast4nodes = read_edges_and_nodes_withoptimIO (argv[1],argv[2],argv[3],argv[4],argv[5],&required_sequence, edgeloader, nodeloader); if (atleast4nodes){ nb_nodes = (int)label.size(); map label_to_node; // Set the label mapping, necessary only to read the edges for (int i = 0; i < nb_nodes; i++) label_to_node[label[i]] = i; // This is a edge-weighted directed graph. The forward and reverse // node in the bidirected de Bruijn graph are split in two nodes. WeightedDigraph G(2 * nb_nodes); for (int i = 0; i < (int)allEdges.size(); i++) { int u = label_to_node[allEdges[i].getFirst()] + (allEdges[i].label[0] == 'F' ? 0 : nb_nodes); int v = label_to_node[allEdges[i].getSecond()] + (allEdges[i].label[1] == 'F' ? 0 : nb_nodes); int cost = strlen(seqs[ label_to_node[allEdges[i].getSecond()] ]) - (k_value - 1); G.adj_list[u].push_back(WeightedEdge(v, cost)); } // We add weights for the nodes only for convenience. All algorithms // work under the assumption of a edge-weighted directed graph. for (int i = 0; i < (int)G.adj_list.size(); i++) G.node_cost[i] = strlen(seqs[i % nb_nodes]) - (k_value - 1); a1Global = LL_MAX - (k_value - 1); a2Global = UL_MAX - (k_value - 1); beta = LL_MIN - (k_value - 1); if (!EXPERIMENTAL_ALG) { listAllBubbles(G, k_value, UL_MAX, LL_MAX, LL_MIN); } else { listAllBubblesUsingPath(G); } } printSummary(stdout, nbBubbles); for (unsigned int i=0; i allPaths; //will contain all paths from s to any t with bounded length == a2 with at most b branching vertices vector explored; //will keep track of the nodes that were already explored on currentPath Path currentPath; ////////////////////////////////////////////////////////////////////////////////////////////// ////////////////////////////////////////////////////////////////////////////////////////////// //helper functions //check if a node is branching or not int isBranching (int node, WeightedDigraph& G) { return (G.outDegree(node) != 1 || inDegree(node, G) != 1) ? 1 : 0; } //check if both paths are vertex-disjoint bool vertexDisjoint (const vector &p1, const vector &p2) { for (int i=1;i<(int)p1.size()-1;i++) { if (find(p2.begin(), p2.end(), p1[i]) != p2.end()) return false; } return true; } //get the number of branching nodes if a new vertex is added to this path int getNbBranchingNodesIfAddAVertex(const Path &path, WeightedDigraph& G) { if (path.nodes.size() <= 1) return 0; return path.branchingNodes + isBranching(path.nodes.back(), G); } ////////////////////////////////////////////////////////////////////////////////////////////// //Recursive function that finds all simple paths from a source s such that the path respects MAX_BRANCHES, beta and a2 void DFSEnum(WeightedDigraph& G, int s) { //add s to the path explored[s]=true; //s is in the current Path //add this path to allPaths if (currentPath.distance >= beta) allPaths.push_back(currentPath); for (int e = 0; e < (int)G.adj_list[s].size(); e++) { //iterates over all edges of s int v = G.adj_list[s][e].node; //check if v should be explored int cost = G.node_cost[s]; int newNbBranchingNodes = getNbBranchingNodesIfAddAVertex(currentPath, G); int newCost = (currentPath.nodes.size() > 1 ? currentPath.distance + cost : 0); if (!explored[v] && //if v is not already in the path newNbBranchingNodes <= MAX_BRANCHES && newCost <= a2Global) { //yes, v should be explored //Configure the currentPath accordingly int oldDistance = currentPath.distance; currentPath.distance = newCost; int oldBranchingNodes = currentPath.branchingNodes; currentPath.branchingNodes = newNbBranchingNodes; currentPath.nodes.push_back(v); //call DFS DFSEnum(G, v); //DesConfigure the currentPath accordingly currentPath.distance = oldDistance; currentPath.branchingNodes = oldBranchingNodes; currentPath.nodes.pop_back(); } } //already explored all paths with this prefix explored[s]=false; //s is not in the currentPath } //list all bubbles by listing all simple paths void findAllBubblesUsingSimpleQueueDFS(WeightedDigraph& G, int source) { //initialize the global variables allPaths.clear(); fill(explored.begin(), explored.end(), false); currentPath.branchingNodes=0; currentPath.distance=0; currentPath.nodes.clear(); //find all paths currentPath.nodes.push_back(source); DFSEnum(G, source); //now list all bubbles map > targetToDistPath; for (vector::iterator it = allPaths.begin(); it != allPaths.end(); ++it) targetToDistPath[it->nodes.back()].push_back(&(*it)); for (map >::iterator targetToDistPathIt = targetToDistPath.begin(); targetToDistPathIt != targetToDistPath.end(); ++targetToDistPathIt) { vector allSTPaths = targetToDistPathIt->second; for (vector::iterator i = allSTPaths.begin(); i != allSTPaths.end(); ++i) { vector::iterator j = i; for (++j; j != allSTPaths.end(); ++j) { Path* lowerPath; Path* upperPath; if ( (*i)->distance < (*j)->distance) { lowerPath = *i; upperPath = *j; } else { lowerPath = *j; upperPath = *i; } if ( (lowerPath->distance <= a1Global || (OUTPUT_SNPS==2 && lowerPath->distance==upperPath->distance) ) && //if we respect LL_MAX (this is for type 0a, 1, 2 and 3) OR we should output all Type0b and both paths have the same length (this is only for type0b) (upperPath->nodes.size() >= 3 || lowerPath->nodes.size() >= 3) && vertexDisjoint(lowerPath->nodes, upperPath->nodes)) { processBubbleFound(upperPath->nodes, lowerPath->nodes, G); } } } } } void listAllBubblesUsingPath(WeightedDigraph& G) { explored = vector(G.adj_list.size(), false); //list all bubbles starting with each node v for (int v = 0; v < (int) G.adj_list.size(); v++) { if (G.outDegree(v) >= 2) //pruning findAllBubblesUsingSimpleQueueDFS(G, v); } } kissplice-2.6.2/modules/BubbleEnumeration.h000066400000000000000000000124461423273700400207430ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #include #include #include #include #include #include #include #include "NGraph.h" #include "Utils.h" #ifndef MOUTH_ENUMERATION_H #define MOUTH_ENUMERATION_H template bool read_edges_and_nodes_withoptimIO(char* filename_info, char* filename_contents_edge, char* filename_contents_node, char* filename_edge, char* filename_node, int *required_sequence, TEdgeFunctor edgefunctor, TNodeFunctor& nodefunctor) { //////////////////////////// //////////////////////////// //////////////////////////// // IO optimization (start) //////////////////////////// int bcc_size, records_per_file, number_of_files_max, file_index; FILE *info_file = fopen(filename_info, "r"); if (info_file == NULL) { fprintf(stderr, "Problem opening %s!\n", filename_info); exit(0); } FILE *contents_file_edge = fopen(filename_contents_edge, "r"); if (contents_file_edge == NULL) { fprintf(stderr, "Problem opening %s!\n", filename_contents_edge); exit(0); } FILE *contents_file_node = fopen(filename_contents_node, "r"); if (contents_file_node == NULL) { fprintf(stderr, "Problem opening %s!\n", filename_contents_node); exit(0); } // read the info file fscanf(info_file, "%d \n",&bcc_size ); fscanf(info_file, "%d \n",&records_per_file ); // find in which file is the required record number_of_files_max = NUMBEROFFILES; if (bcc_sizenumber_of_files_max) file_index = number_of_files_max; if ( (*required_sequence <=0) || (*required_sequence > bcc_size) ) { fprintf(stderr, "Problem opening sequence %d in edge/node files !\n", *required_sequence); exit(0); } // filenames char total_edge_fname[1024]; sprintf( total_edge_fname, "%s_%d",filename_edge,file_index ); char total_node_fname[1024]; sprintf( total_node_fname, "%s_%d",filename_node,file_index ); //////////////////////////// // IO optimization (end) //////////////////////////// //////////////////////////// //////////////////////////// FILE* edge_file = open_file(total_edge_fname); FILE* node_file = open_file(total_node_fname); bool atleast4nodes = read_node_noncontigous_file_withoptimIO( contents_file_node, node_file, required_sequence, &file_index, nodefunctor); if (atleast4nodes){ read_edge_file_withoptimIO( contents_file_edge, edge_file, required_sequence, &file_index, edgefunctor ); } fclose( contents_file_edge ); fclose( contents_file_node ); fclose( edge_file ); fclose( node_file ); return atleast4nodes; } //Type used to represent a path. //It keeps track of the length (distance) of the path, its number of branching nodes and its nodes struct Path { int distance; int branchingNodes; vector nodes; Path():distance(0), branchingNodes(0), nodes(){} Path (int distance, int branchingNodes, const vector &nodes) : distance(distance), branchingNodes(branchingNodes), nodes(nodes){} }; #endif /* MOUTH_ENUMERATION_H */ kissplice-2.6.2/modules/CEdge.h000066400000000000000000000136651423273700400163140ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #ifndef EDGE_H #define EDGE_H // =========================================================================== // Include Libraries // =========================================================================== #include #include // =========================================================================== // Include Project Files // =========================================================================== // =========================================================================== // Class declarations // =========================================================================== //! Edge class /*! * \brief Class meant to be used by CGraph (compact graph) to represent edges. * * It encapsulates a pair of int (u -> v edge), each int (u or v) being a node id */ class CEdge { public : // ======================================================================= // Enums // ======================================================================= // ======================================================================= // Constructors // ======================================================================= CEdge() = default; CEdge( const CEdge& ) = default; CEdge( int f, int s ) : m_pair( f, s ) {} // ======================================================================= // Destructor // ======================================================================= // ======================================================================= // Accessors: getters // ======================================================================= inline int getFirst() const { return m_pair.first; } inline int getSecond() const { return m_pair.second; } // ======================================================================= // Accessors: setters // ======================================================================= // ======================================================================= // Operators // ======================================================================= bool operator==( const CEdge& that ) const { return m_pair == that.m_pair; } bool operator!=( const CEdge& that ) const { return m_pair != that.m_pair; } bool operator<( const CEdge& that ) const { return m_pair < that.m_pair; } // ======================================================================= // Public Methods // ======================================================================= CEdge& swap_ends() { std::swap(m_pair.first, m_pair.second); return *this; } // ======================================================================= // Public Attributes // ======================================================================= protected : std::pair m_pair; // ======================================================================= // Forbidden Constructors // ======================================================================= // ======================================================================= // Protected Methods // ======================================================================= // ======================================================================= // Protected Attributes // ======================================================================= }; #endif // EDGE_H kissplice-2.6.2/modules/CGraph.cpp000066400000000000000000000160301423273700400170310ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ // =========================================================================== // Include Libraries // =========================================================================== #include #include // =========================================================================== // Include Project Files // =========================================================================== #include "CGraph.h" // =========================================================================== // Declare Used Namespaces // =========================================================================== using namespace std; //############################################################################ // # // Class CGraph # // # //############################################################################ // =========================================================================== // Constructors // =========================================================================== CGraph::CGraph(int nbNodes, vector& allEdges, int kValue) { k_value = kValue; adj_list.reserve(nbNodes); adj_list_sz.reserve(nbNodes); int j = 0; int offset = 0; for ( int i = 0 ; i < nbNodes ; i++ ) { // Count the number of edges (they are ordered in all_edges) of node i j = 0; while ( offset + j < (int) allEdges.size() && allEdges[offset + j].getFirst() == i ) { j++; } // ... adj_list.push_back(j != 0 ? new int[j] : NULL); adj_list_sz.push_back(0); offset += j; } // ... for ( int i = 0 ; i < (int)allEdges.size() ; i++ ) { insert_unique_edge_dir( allEdges[i].getFirst(), allEdges[i].getSecond() ); } } // =========================================================================== // Public Methods // =========================================================================== void CGraph::insert_unique_edge_dir(int u, int v) { int i; int* neighbours = adj_list[u]; int nb_neighbours = adj_list_sz[u]; // Find corresponding neighbour (if it exists) for ( i = 0 ; i < nb_neighbours ; i++, neighbours++ ) { if ( *neighbours == v ) { break; } } // If not found, add the missing edge if ( i == nb_neighbours ) { *neighbours = v; adj_list_sz[u]++; } } void CGraph::insert_unique_edge( int u, int v ) { insert_unique_edge_dir(u,v); insert_unique_edge_dir(v,u); } void CGraph::destroy_adj_list(void) { for (int i = 0; i < (int)adj_list.size(); i++) delete[] adj_list[i]; vector().swap(adj_list); } void CGraph::destroy(void) { destroy_adj_list(); vector().swap(adj_list_sz); } //================================================================ // Reading functions //================================================================ int count_nb_lines( FILE* file ) { int ch, number_of_lines = 0; while (EOF != (ch=getc(file))) if ('\n' == ch) number_of_lines++; // Set the cursor back to the begining of the file. rewind(file); // Don't care if the last line has a '\n' or not. We over-estimate it. return number_of_lines + 1; } void read_node_file( FILE* node_file, vector& seqs, int k_val ) { char* buffer = new char[100 * MAX]; char* seq; seqs.reserve(count_nb_lines(node_file)); while ( fgets(buffer, 100 * MAX, node_file) != NULL ) { char* p; if (strlen(buffer) == 100 * MAX) { p = strtok(buffer, "\t\n"); fprintf(stdout, "ERROR: node %s with sequence larger than %d!", p, 100 * MAX); exit(0); } // Node label p = strtok( buffer, "\t\n" ); // Node seq p = strtok( NULL, "\t\n" ); seq = new char[strlen(p) + 1]; strcpy( seq, p ); seqs.push_back( seq ); //~ printf( "pushed %s (0x%x)|\n", seq, seq ); //~ getchar(); //~ delete [] seq; } delete [] buffer; } void read_edge_file( FILE *edge_file, vector& edges ) { char* buffer = new char[100 * MAX]; char* u = new char[MAX]; char* v = new char[MAX]; char* label = new char[MAX]; edges.reserve(count_nb_lines(edge_file)); while ( fgets(buffer, 100 * MAX, edge_file) != NULL ) { char* p; // outgoing node p = strtok( buffer, "\t\n" ); strcpy( u, p ); // incoming node p = strtok( NULL, "\t\n" ); strcpy( v, p ); // edge label p = strtok( NULL, "\t\n" ); strcpy(label, p); edges.push_back( LabelledCEdge( atoi(u), atoi(v), label ) ); } sort( edges.begin(), edges.end() ); delete [] buffer; delete [] u; delete [] v; delete [] label; } kissplice-2.6.2/modules/CGraph.h000066400000000000000000000270701423273700400165040ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #ifndef COMPRESSED_GRAPH_H #define COMPRESSED_GRAPH_H // =========================================================================== // Include Libraries // =========================================================================== #include #include #include #include #include #include // =========================================================================== // Include Project Files // =========================================================================== #include "CEdge.h" #include "LabelledCEdge.h" // =========================================================================== // Class declarations // =========================================================================== // =========================================================================== // Declare Used Namespaces // =========================================================================== using namespace std; //! A class representing the compacted De-Bruijn graphs /*!\ * \brief Its light structure is used for the detection of bi-connected * component in the graph. */ #define MAX 1024 class CGraph { public: // ======================================================================= // Constructors // ======================================================================= /*! * \brief CGraph constructor. * Initializes the attributes according to the node numbers * Creates directly the CGraph object with its edges from allEdges */ CGraph( int nbNodes, vector & allEdges, int kValue ); // ======================================================================= // Public Attributes // ======================================================================= int k_value; // ======================================================================= // Public Methods // ======================================================================= inline int *get_adj_list(int node) const; inline int get_adj_list_sz(int node) const; inline int get_n_nodes(void) const; void insert_unique_edge_dir(int u, int v); void insert_unique_edge(int u, int v); void destroy_adj_list(void); void destroy(void); protected: // ======================================================================= // Forbidden Constructors // ======================================================================= CGraph(const CGraph &) = delete; CGraph() = delete; // ======================================================================= // Protected Methods // ======================================================================= // ======================================================================= // Protected Attributes // ======================================================================= // NOTA: The neighborhood size is always <= 8 //! Adjacency list /*! * adj_list is a vector of int tables containing the IDs of "outgoing neighbours" * i.e. neighbours v having a u -> v edge (u being the present node). * adj_list_sz contains the corresponding sizes */ vector adj_list; vector adj_list_sz; }; // =========================================================================== // Inline functions' definition // =========================================================================== int *CGraph::get_adj_list(int node) const { return adj_list[node]; } int CGraph::get_adj_list_sz(int node) const { return adj_list_sz[node]; } int CGraph::get_n_nodes(void) const { return (int) adj_list.size(); } // =========================================================================== // Other functions // =========================================================================== void read_node_file(FILE *node_file, vector& seqs, int k_val); void read_edge_file(FILE *edge_file, vector& edges); // Functor that load the bcc nodes in memory class NodeLoader { private: vector& _seqs; vector& _label; public: NodeLoader(vector& seqs, vector& label) : _seqs(seqs), _label(label) {} void operator()(FILE* node_file, int written_lines){ char* buffer = new char[100 * MAX]; for (int i = 0; i < written_lines ; i++) { char* p; fgets(buffer, 100 * MAX, node_file); if (strlen(buffer) == 100 * MAX) { p = strtok(buffer, "\t\n"); fprintf(stdout, "ERROR: node %s with sequence larger than %d!", p, 100 * MAX); exit(0); } // Node label p = strtok( buffer, "\t\n" ); _label.push_back(atoi(p)); // Node seq p = strtok( NULL, "\t\n" ); char* seq = new char[strlen(p) + 1]; strcpy( seq, p ); // Node seq_r p = strtok( NULL, "\t\n" ); _seqs.push_back( seq ); } delete [] buffer; } }; // return false if nbnodes<4 // required a functor to acts on the file lines template bool read_node_noncontigous_file_withoptimIO( FILE *contents_file_node, FILE* node_file, int *required_sequence, int *file_index, TFunctor& functor ) { char* buffer = new char[100 * MAX]; //////////////////////////// //////////////////////////// //////////////////////////// // IO optimization (start) //////////////////////////// int written_lines_before, written_lines_for_record, written_lines; //normally would skip directly to the record, but for the time being use the formatted file for (int i = 0; i < *required_sequence+*file_index-2 ; i++) fgets(buffer, 100 * MAX, contents_file_node); fscanf(contents_file_node, "%d \n",&written_lines_before ); fscanf(contents_file_node, "%d \n",&written_lines_for_record ); // if a graph node was not written to disk if ( written_lines_before == written_lines_for_record) { fprintf(stderr, "The required record %d does not exist in the bcc graph !\n", *required_sequence); exit(0); } // where to search the record written_lines = written_lines_for_record - written_lines_before; bool atleast4nodes = false; if (written_lines>=4){ //Less than 4 nodes, cannot contain a bubble! atleast4nodes = true; //normally would skip directly to the record, but for the time being use the formatted file for (int i = 0; i < written_lines_before ; i++) fgets(buffer, 100 * MAX, node_file); //////////////////////////// // IO optimization (end) //////////////////////////// //////////////////////////// //////////////////////////// functor(node_file, written_lines); } delete [] buffer; return atleast4nodes; } // Functor that load the bcc edges in memory class EdgeLoader { private: vector& _edges; public: EdgeLoader(vector& edges) : _edges(edges) {} void operator()(FILE* edge_file, int written_lines){ if (written_lines>0){ char* buffer = new char[100 * MAX]; char* u = new char[MAX]; char* v = new char[MAX]; char* label = new char[MAX]; _edges.reserve(written_lines);//count_nb_lines(edge_file)); for (int i = 0; i < written_lines ; i++) { char* p; fgets(buffer, 100 * MAX, edge_file); // outgoing node p = strtok( buffer, "\t\n" ); strcpy( u, p ); // incoming node p = strtok( NULL, "\t\n" ); strcpy( v, p ); // edge label p = strtok( NULL, "\t\n" ); strcpy(label, p); _edges.push_back( LabelledCEdge( atoi(u), atoi(v), label ) ); } sort( _edges.begin(), _edges.end() ); delete [] u; delete [] v; delete [] label; delete [] buffer; } } }; // required a functor to acts on the file lines template void read_edge_file_withoptimIO( FILE *contents_file_edge, FILE *edge_file, int *required_sequence, int *file_index, TFunctor& functor ) { char* buffer = new char[100 * MAX]; //////////////////////////// //////////////////////////// //////////////////////////// // IO optimization (start) // [replicate as above] //////////////////////////// int written_lines_before, written_lines_for_record, written_lines; //normally would skip directly to the record, but for the time being use the formatted file for (int i = 0; i < *required_sequence+*file_index-2 ; i++) fgets(buffer, 100 * MAX, contents_file_edge); fscanf(contents_file_edge, "%d \n",&written_lines_before ); fscanf(contents_file_edge, "%d \n",&written_lines_for_record ); // if a graph edge was not written to disk if ( written_lines_before == written_lines_for_record) { fprintf(stderr, "The required record %d does not exist in the bcc graph !\n", *required_sequence); exit(0); } // where to search the record written_lines = written_lines_for_record - written_lines_before; //normally would skip directly to the record, but for the time being use the formatted file for (int i = 0; i < written_lines_before ; i++) fgets(buffer, 100 * MAX, edge_file); //////////////////////////// // IO optimization (end) //////////////////////////// //////////////////////////// //////////////////////////// functor(edge_file, written_lines); delete [] buffer; } #endif // COMPRESSED_GRAPH_H kissplice-2.6.2/modules/CMakeLists.txt000066400000000000000000000026321423273700400177240ustar00rootroot00000000000000add_executable(ks_clean_duplicates CleanDuplicates.cpp CycleCompression.cpp NGraph.cpp CGraph.cpp LabelledCEdge.cpp Utils.cpp) set_property(TARGET ks_clean_duplicates PROPERTY RUNTIME_OUTPUT_DIRECTORY "${BUILD_INTERNAL_BINDIR}") install(TARGETS ks_clean_duplicates RUNTIME DESTINATION "${RELATIVE_INTERNAL_BINDIR}") add_executable(ks_bubble_enumeration BubbleEnumeration.cpp CycleCompression.cpp NGraph.cpp CGraph.cpp LabelledCEdge.cpp Utils.cpp) set_property(TARGET ks_bubble_enumeration PROPERTY RUNTIME_OUTPUT_DIRECTORY "${BUILD_INTERNAL_BINDIR}") install(TARGETS ks_bubble_enumeration RUNTIME DESTINATION "${RELATIVE_INTERNAL_BINDIR}") add_executable(ks_run_modules run.cpp CycleCompression.cpp NGraph.cpp CGraph.cpp LabelledCEdge.cpp SplitBcc.cpp Utils.cpp) set_property(TARGET ks_run_modules PROPERTY RUNTIME_OUTPUT_DIRECTORY "${BUILD_INTERNAL_BINDIR}") install(TARGETS ks_run_modules RUNTIME DESTINATION "${RELATIVE_INTERNAL_BINDIR}") add_executable(ks_error_removal ErrorRemoval.cpp LabelledCEdge.cpp Utils.cpp) set_property(TARGET ks_error_removal PROPERTY RUNTIME_OUTPUT_DIRECTORY "${BUILD_INTERNAL_BINDIR}") install(TARGETS ks_error_removal RUNTIME DESTINATION "${RELATIVE_INTERNAL_BINDIR}") add_executable(ks_print_bcc PrintBcc.cpp Utils.cpp) set_property(TARGET ks_print_bcc PROPERTY RUNTIME_OUTPUT_DIRECTORY "${BUILD_INTERNAL_BINDIR}") install(TARGETS ks_print_bcc RUNTIME DESTINATION "${RELATIVE_INTERNAL_BINDIR}")kissplice-2.6.2/modules/CleanDuplicates.cpp000066400000000000000000000100021423273700400207160ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #include #include #include #include #include #include "Utils.h" #include "NGraph.h" using namespace std; struct block { string l[4]; }; bool comp_block(block a, block b) { string a_up = a.l[1]; string a_low = a.l[3]; string b_up = b.l[1]; string b_low = b.l[3]; // upper path is the lexicograficaly smaller, canonical form. if (a_low < a_up) swap(a_low, a_up); if (b_low < b_up) swap(b_low, b_up); return (a_up < b_up) || (a_up == b_up && a_low < b_low); } bool eq(block a, block b) { string a_up = a.l[1], a_low = a.l[3], b_up = b.l[1], b_low = b.l[3]; // upper path is the lexicograficaly smaller, canonical form. if (a_low < a_up) swap(a_low, a_up); if (b_low < b_up) swap(b_low, b_up); return a_up == b_up && a_low == b_low; } block reverse_complement(block &b) { block res; res.l[1] = reverse_complement(b.l[1]); res.l[3] = reverse_complement(b.l[3]); res.l[0] = b.l[0]; res.l[2] = b.l[2]; return res; } int main(int argc, char **argv) { ifstream bubbles_file; bubbles_file.open(argv[1]); string line; int nlines = 0; block cur; vector all_blocks; while (bubbles_file.good()) { getline(bubbles_file, line); nlines++; cur.l[(nlines-1) % 4] = line; if (nlines % 4 == 0) { // convert the sequences to a canonical form, lexicografically smaller (forward and reverse) if (comp_block(reverse_complement(cur), cur)) cur = reverse_complement(cur); all_blocks.push_back(cur); } } sort(all_blocks.begin(), all_blocks.end(), comp_block); vector::iterator end = unique(all_blocks.begin(), all_blocks.end(), eq); vector::iterator it; for (it = all_blocks.begin(); it != end; it++) for (int j = 0; j < 4; j++) cout << it->l[j] << "\n"; bubbles_file.close(); return 0; } kissplice-2.6.2/modules/ClusterBubbles.cpp000066400000000000000000000141321423273700400206060ustar00rootroot00000000000000#include #include #include #include #include #include #include #include using namespace std; #define MIN(a,b) ((a) < (b) ? (a) : (b)) #define MIN3(a,b,c) ((a) < (b) ? ((a) < (c) ? (a) : (c)) : ((b) < (c) ? (b) : (c))) struct block { string l[4], bcc; double rank; }; static char complement(char b) { switch(b) { case 'A': return 'T'; case 'T': return 'A'; case 'G': return 'C'; case 'C': return 'G'; case 'a': return 't'; case 't': return 'a'; case 'g': return 'c'; case 'c': return 'g'; case 'N': return 'N'; case '*': return '*'; } return '?'; } string reverse_complement(string seq) { string s(seq.begin(),seq.end()); string::iterator pos; for (pos = s.begin(); pos != s.end(); ++pos) { // cout << *pos; } // cout << endl; reverse(s.begin(), s.end()); for(pos=s.begin();pos!=s.end();++pos) *pos=complement(*pos); return s; } bool comp_lower(block a, block b) { return a.l[3] < b.l[3]; } bool comp_bcc(block a, block b) { return a.bcc < b.bcc; } block reverse_complement(block &b) { block res; res.l[1] = reverse_complement(b.l[1]); res.l[3] = reverse_complement(b.l[3]); res.l[0] = b.l[0]; res.l[2] = b.l[2]; return res; } string extract_bcc(block &b) { int sep = b.l[0].find_first_of("|"); return b.l[0].substr(0, sep); } double extract_rank(block &b) { int sep = b.l[0].find_last_of("|"); string rank = b.l[0].substr(sep+1); sep = rank.find_first_of("_"); float res = atof(rank.substr(sep+1).c_str()); /* trick to test is res = nan */ if (res != res) res = 0; return res; } void print_block(FILE *stream, block &b) { for (int i = 0; i < 4; i++) fprintf(stream, "%s\n", b.l[i].c_str()); } map > adj; vector all_blocks; int max_rank, max_ind; void dfs(int node, map &visited) { visited[node] = true; if (all_blocks[node].rank >= max_rank) { max_rank = all_blocks[node].rank; max_ind = node; } for (int i = 0; i < (int)adj[node].size(); i++) { int v = adj[node][i]; if (!visited[v]) dfs(v, visited); } } int nprinted = 0; void print_clusters(FILE *stream) { map visited; map >::iterator it; for (it = adj.begin(); it != adj.end(); it++) visited[it->first] = false; for (it = adj.begin(); it != adj.end(); it++) if (!visited[it->first]) { max_rank = max_ind = -1; dfs(it->first, visited); if (max_ind != -1) { nprinted++; print_block(stream, all_blocks[it->first]); } } } int comp_base(const void *a, const void *b) { return *(char *)a != *(char *)b; } int edit_distance(const void *s1, size_t l1, const void *s2, size_t l2, size_t nmemb, int (*comp)(const void*, const void*)) { unsigned int i; unsigned int j; size_t len = (l1 + 1) * (l2 + 1); char *p1; char *p2; unsigned int d1; unsigned int d2; unsigned int d3; unsigned int *d; unsigned int *dp; unsigned int res; if (l1 == 0) return l2; else if (l2 == 0) return l1; d = (unsigned int*)malloc(len * sizeof(unsigned int)); *d = 0; for(i = 1, dp = d + l2 + 1; i < l1 + 1; ++i, dp += l2 + 1) { *dp = (unsigned) i; } for(j = 1, dp = d + 1; j < l2 + 1; ++j, ++dp) { *dp = (unsigned) j; } for(i = 1, p1 = (char*) s1, dp = d + l2 + 2; i < l1 + 1; ++i, p1 += nmemb, ++dp) { for(j = 1, p2 = (char*) s2; j < l2 + 1; ++j, p2 += nmemb, ++dp) { if(!comp(p1, p2)) { *dp = *(dp - l2 - 2);// same base } else { d1 = *(dp - 1) + 1;// insertion d2 = *(dp - l2 - 1) + 1;// deletion d3 = *(dp - l2 - 2) + 1;// substitution *dp = MIN3(d1, d2, d3); } } } res = *(dp - 2); dp = NULL; free(d); return res; } int min_dist = 1; bool has_edge(int i, int j) { // lower path should be equal if (all_blocks[i].l[3] != all_blocks[j].l[3]) return false; if (abs((int)all_blocks[i].l[1].size() - (int)all_blocks[j].l[1].size()) > min_dist) return false; int ed = edit_distance(all_blocks[i].l[1].c_str(), all_blocks[i].l[1].size(), all_blocks[j].l[1].c_str(), all_blocks[j].l[1].size(), sizeof(char), comp_base); return (ed <= min_dist); } int main(int argc, char **argv) { ifstream bubbles_file; bubbles_file.open(argv[1]); if (argc < 3) { fprintf(stderr, "./cluster input.fasta output.fasta [min_dist]\n"); exit(0); } if (argc == 4) min_dist = atoi(argv[3]); string line; int nlines = 0; block cur; while (bubbles_file.good()) { getline(bubbles_file, line); nlines++; cur.l[(nlines-1) % 4] = line; if (nlines % 4 == 0) { // convert the sequences to a canonical form, lexicografically smaller (forward and reverse) if (comp_lower(reverse_complement(cur), cur)) cur = reverse_complement(cur); // extract the bcc number cur.bcc = extract_bcc(cur); cur.rank = extract_rank(cur); all_blocks.push_back(cur); } } FILE *output = fopen(argv[2], "w"); if (output == NULL) { fprintf(stderr, "Problem opening %s\n", argv[2]); exit(0); } // sort by BCC, all bubbles from the same BCC are adjacent sort(all_blocks.begin(), all_blocks.end(), comp_bcc); for (int i = 0; i < (int)all_blocks.size(); i++) { map >::iterator it; adj[i] = vector(); for (it = adj.begin(); it != adj.end(); it++) if (has_edge(i, it->first)) { adj[i].push_back(it->first); adj[it->first].push_back(i); } // finished the bubbles for a given BCC if ((i+1) == (int)all_blocks.size() || all_blocks[i].bcc != all_blocks[i+1].bcc) { print_clusters(output); map >().swap(adj); } } fclose(output); fprintf(stdout, "%d out of %d bubbles were outputed\n", nprinted, (int)all_blocks.size()); //fprintf(stderr, "nprinted = %d, total = %d, max_nodes = %d, max_edges = %d\n", nprinted, (int)all_blocks.size(), max_sz, max_edges); bubbles_file.close(); return 0; } kissplice-2.6.2/modules/CycleCompression.cpp000066400000000000000000000106621423273700400211530ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #include #include #include #include #include #include "debug.h" #include "NGraph.h" #include "Utils.h" #include "CycleCompression.h" #define MAX 1024 //MAX2 and MIN2 never used? TODO: Remove? #define MAX2(a,b) ((a) > (b) ? (a) : (b)) #define MIN2(a,b) ((a) < (b) ? (a) : (b)) using namespace std; idx_dir reverse_dir(idx_dir node) { return make_pair(node.first, reverse_dir(node.second)); } bool is_same_node( const idx_dir u, const idx_dir v) { return (v.first == u.first && v.second == u.second); } /*! * \brief Merge sequence information between two nodes and print it in the snp file * with bbc number, cycle and length of upper and lower path. * Returns the merged sequence * \param seq1 sequence of the upper path * \param seq2 sequence of the lower path * \param snp_log_file the file to be printing the two paths * \param bccid id of the bi-connected component being treated */ string merge_sequences( const string seq1, const string seq2 ) { string merged; for (int i = 0 ; i < (int)seq1.size(); i++) merged += (seq1[i] == seq2[i] ? seq1[i] : 'N'); return merged; } void output_sequences( string seq1, string seq2, FILE * snp_log_file, const int bccid, const int cycleNum, const int contextL, const int contextR, const int kValue) { int u_len = (int)seq1.size(); int l_len = (int)seq2.size(); if( contextL != 0 || contextR !=0 ) { seq1 = toLowerContext(seq1, contextL, contextR); seq2 = toLowerContext(seq2, contextL, contextR); } if (u_len- (contextL + contextR) > 2* kValue +1) // multiple SNP { fprintf(snp_log_file, ">bcc_%d|Cycle_%d|Type_0b|upper_path_Length_%d\n", bccid, cycleNum, u_len- (contextL + contextR)); fprintf(snp_log_file, "%s\n", seq1.c_str()); fprintf(snp_log_file, ">bcc_%d|Cycle_%d|Type_0b|lower_path_Length_%d\n", bccid, cycleNum, l_len- (contextL + contextR)); fprintf(snp_log_file, "%s\n", seq2.c_str()); } else // single SNP { fprintf(snp_log_file, ">bcc_%d|Cycle_%d|Type_0a|upper_path_Length_%d\n", bccid, cycleNum, u_len- (contextL + contextR)); fprintf(snp_log_file, "%s\n", seq1.c_str()); fprintf(snp_log_file, ">bcc_%d|Cycle_%d|Type_0a|lower_path_Length_%d\n", bccid, cycleNum, l_len- (contextL + contextR)); fprintf(snp_log_file, "%s\n", seq2.c_str()); } } kissplice-2.6.2/modules/CycleCompression.h000066400000000000000000000052321423273700400206150ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #include #include #include #include "NGraph.h" #include "Utils.h" #ifndef CYCLE_COMPRESSION_H #define CYCLE_COMPRESSION_H // Auxiliary functions string merge_sequences( const string seq1, const string seq2); void output_sequences( string seq1, string seq2, FILE * snp_log_file, const int bccid, const int cycleNumconst, int contextL, const int contextR, const int kValue); bool is_same_node(const idx_dir u, const idx_dir v); idx_dir reverse_dir(idx_dir node); #endif /* CYCLE_COMPRESSION_H */ kissplice-2.6.2/modules/EntropyFilter.cpp000066400000000000000000000100161423273700400204710ustar00rootroot00000000000000#include #include #include #include #include #include #include #include using namespace std; #define MIN(a,b) ((a) < (b) ? (a) : (b)) #define MIN3(a,b,c) ((a) < (b) ? ((a) < (c) ? (a) : (c)) : ((b) < (c) ? (b) : (c))) struct block { string l[4], bcc; double rank; }; static char complement(char b) { switch(b) { case 'A': return 'T'; case 'T': return 'A'; case 'G': return 'C'; case 'C': return 'G'; case 'a': return 't'; case 't': return 'a'; case 'g': return 'c'; case 'c': return 'g'; case 'N': return 'N'; case '*': return '*'; } return '?'; } string reverse_complement(string seq) { string s(seq.begin(),seq.end()); string::iterator pos; for (pos = s.begin(); pos != s.end(); ++pos) { // cout << *pos; } // cout << endl; reverse(s.begin(), s.end()); for(pos=s.begin();pos!=s.end();++pos) *pos=complement(*pos); return s; } block reverse_complement(block &b) { block res; res.l[1] = reverse_complement(b.l[1]); res.l[3] = reverse_complement(b.l[3]); res.l[0] = b.l[0]; res.l[2] = b.l[2]; return res; } void print_block(FILE *stream, block &b) { for (int i = 0; i < 4; i++) fprintf(stream, "%s\n", b.l[i].c_str()); } int nucToNumber(const char& nuc) { switch (nuc) { case 'A': return 0; case 'C': return 1; case 'G': return 2; case 'T': return 3; default: return 4; } } static double MinusPlogP(double value, double base) { return -1 * value * log(value)/log(base); } bool valid_nuc(char nuc) { return nuc != 'N' && nuc != '*' && nuc != '?'; } double entropy3(const string& window_nucs) { size_t window_length = window_nucs.length(); vector kmer_counts(444, 0); float subseqs = 0; for (size_t i = 0; i < window_length - 3; ++i) { char nuc1 = window_nucs.at(i); char nuc2 = window_nucs.at(i+1); char nuc3 = window_nucs.at(i+2); if (valid_nuc(nuc1) && valid_nuc(nuc2) && valid_nuc(nuc3)) { kmer_counts[nucToNumber(nuc1) + nucToNumber(nuc2)*10 + nucToNumber(nuc3)*10] += 1; subseqs+= 1; } } float entropy = 0, base = 64; if (window_length < 66) base = (float)window_length; for (size_t i = 0; i < kmer_counts.size(); ++i) { int count = kmer_counts.at(i); if (count !=0) { float p = static_cast(count)/subseqs; entropy += MinusPlogP(p, base); } } return entropy; } bool comp_bcc(block a, block b) { return a.bcc < b.bcc; } string extract_bcc(block &b) { int sep = b.l[0].find_first_of("|"); return b.l[0].substr(0, sep); } int main(int argc, char **argv) { ifstream bubbles_file; bubbles_file.open(argv[1]); double threshold = 0.70; if (argc < 3) { fprintf(stderr, "./entropy_filter input.fasta output.fasta [threshold]\n"); exit(0); } if (argc == 4) threshold = atof(argv[3]); fprintf(stderr, "%lf\n", threshold); string line; int nlines = 0; block cur; map bcc_count; vector all_blocks; FILE *output = fopen(argv[2], "w"), *filtered = fopen("removed.fa", "w"); if (output == NULL) { fprintf(stderr, "Problem opening %s\n", argv[2]); exit(0); } int nprinted = 0; while (bubbles_file.good()) { getline(bubbles_file, line); nlines++; cur.l[(nlines-1) % 4] = line; if (nlines % 4 == 0) { cur.bcc = extract_bcc(cur); all_blocks.push_back(cur); if (bcc_count.find(cur.bcc) == bcc_count.end()) bcc_count[cur.bcc] = 0; bcc_count[cur.bcc] = bcc_count[cur.bcc] + 1; } } for (int i = 0; i < (int)all_blocks.size(); i++) if (entropy3(all_blocks[i].l[1]) > threshold || bcc_count[all_blocks[i].bcc] <= 10) { nprinted++; print_block(output, all_blocks[i]); } else { print_block(filtered, all_blocks[i]); } fclose(output); fclose(filtered); bubbles_file.close(); fprintf(stdout, "%d out of %d bubbles were outputed\n", nprinted, (int)all_blocks.size()); return 0; } kissplice-2.6.2/modules/ErrorRemoval.cpp000066400000000000000000000143271423273700400203130ustar00rootroot00000000000000#include #include #include #include #include #include "Utils.h" #include "LabelledCEdge.h" #include #include #define MAX 1024 using namespace std; struct NodeSeq { char *seq; int node; NodeSeq(char *initSeq, int initNode) : seq(initSeq), node(initNode) { } bool operator== (const NodeSeq& rhs) const { return (strcmp(seq, rhs.seq) == 0); } bool operator< (const NodeSeq& rhs) const { return (strcmp(seq, rhs.seq) < 0); } }; int count_nb_lines( FILE* file ) { int ch, number_of_lines = 0; while (EOF != (ch=getc(file))) if ('\n' == ch) number_of_lines++; // Set the cursor back to the begining of the file. rewind(file); // Don't care if the last line has a '\n' or not. We over-estimate it. return number_of_lines + 1; } void readEdgeFile( char* nodes_fname, vector& edges ) { FILE *edge_file = open_file(nodes_fname); char* buffer = new char[100 * MAX]; char* u = new char[MAX]; char* v = new char[MAX]; char* label = new char[MAX]; edges.reserve(count_nb_lines(edge_file)); while ( fgets(buffer, 100 * MAX, edge_file) != NULL ) { char* p; // outgoing node p = strtok( buffer, "\t\n" ); strcpy( u, p ); // incoming node p = strtok( NULL, "\t\n" ); strcpy( v, p ); // edge label p = strtok( NULL, "\t\n" ); strcpy(label, p); edges.push_back( LabelledCEdge( atoi(u), atoi(v), label ) ); } sort( edges.begin(), edges.end() ); delete [] buffer; delete [] u; delete [] v; delete [] label; fclose(edge_file); } // rev_seq needs to be global! let's make sure we allocate only once char *rev_seq = NULL; char *revComp(char *seq) { int size = strlen(seq); if (rev_seq == NULL) rev_seq = new char[size + 1]; for (int i = size - 1; i >= 0; i--) rev_seq[(size - 1) - i] = complement(seq[i]); rev_seq[size] = '\0'; return rev_seq; } int findNode(char *query, vector& nodes) { vector::iterator low, low_r; char *query_r = revComp(query); // The value of "x" in NodeSeq(query, x) doesn't matter low = lower_bound(nodes.begin(), nodes.end(), NodeSeq(query, 0)); low_r = lower_bound(nodes.begin(), nodes.end(), NodeSeq(query_r, 0)); if (low != nodes.end() && *low == NodeSeq(query, 0)) return low->node; if (low_r != nodes.end() && *low_r == NodeSeq(query_r, 0)) return low_r->node; return -1; } void readAbundanceFile (char* abundanceFileName, vector &abundances) { ifstream abundanceFile(abundanceFileName); if (!abundanceFile) { cerr << "Error opening " << abundanceFileName << endl; exit(1); } double abundance; while (abundanceFile >> abundance) abundances.push_back(abundance); abundanceFile.close(); } int findEdge(vector& allEdges, LabelledCEdge e) { vector::iterator low; low = lower_bound(allEdges.begin(), allEdges.end(), e); if (low != allEdges.end()) return low - allEdges.begin(); fprintf(stderr, "inconsistent graph!"); return -1; } // Move this to CEdge LabelledCEdge reverse(LabelledCEdge e) { char *label = new char[3]; label[0] = e.label[1] == 'F' ? 'R' : 'F'; label[1] = e.label[0] == 'F' ? 'R' : 'F'; label[2] = '\0'; return LabelledCEdge(e.getSecond(), e.getFirst(), label); } void errorRemoval(vector& allEdges, int nbNodes, double cutoff, vector& removed, const vector &abundances) { int offset = 0; for ( int src = 0 ; src < nbNodes; src++ ) { // Count the number of edges (they are ordered in all_edges) of node src int size = 0; while (offset + size < (int) allEdges.size() && allEdges[offset + size].getFirst() == src) size++; // Compute the sum of coverage for each direction double sum_F = 0, sum_R = 0; for (int k = 0; k < size; k++) { if (!removed[offset + k]) { int target = allEdges[offset + k].getSecond(); if (allEdges[offset + k].label[0] == 'F') sum_F += abundances[target]; if (allEdges[offset + k].label[0] == 'R') sum_R += abundances[target]; } } // Remove the edges with relative coverage below cutoff for (int k = 0; k < size; k++) { if (!removed[offset + k]) { int target = allEdges[offset + k].getSecond(); double count = abundances[target]; double ratio; ratio = count / sum_F; if (allEdges[offset + k].label[0] == 'F' && ratio < cutoff) { removed[offset + k] = true; removed[findEdge(allEdges, reverse(allEdges[offset + k]))] = true; } ratio = count / sum_R; if (allEdges[offset + k].label[0] == 'R' && ratio < cutoff) { removed[offset + k] = true; removed[findEdge(allEdges, reverse(allEdges[offset + k]))] = true; } } } offset += size; } } int main( int argc, char** argv ) { string base_name = "graph"; if ( argc < 4 ) { fprintf( stderr, "Wrong number of arguments!\n" ); fprintf( stderr, "Usage: ./error_removal edge_file unitig_abundance_file cutoff [base_name]\n" ); return 0; } if ( argc == 5 ) { base_name = argv[4]; } double cutoff = atof( argv[3] ); // Read edge file vector allEdges; readEdgeFile(argv[1], allEdges ); // Read abundance file vector abundances; abundances.reserve(allEdges.size()); //over-estimating, but it is ok readAbundanceFile(argv[2], abundances); //read the abundances vector removed(allEdges.size(), false); errorRemoval(allEdges, (int)abundances.size(), cutoff, removed, abundances); int nb_removed = 0; FILE *output = fopen((base_name + ".edges").c_str(), "w"); FILE *removed_out = fopen((base_name + "_removed.edges").c_str(), "w"); for (int i = 0 ; i < (int)allEdges.size(); i++) if (!removed[i]) fprintf(output, "%d\t%d\t%s\n", allEdges[i].getFirst(), allEdges[i].getSecond(), allEdges[i].label); else { fprintf(removed_out, "%d\t%d\t%s\n", allEdges[i].getFirst(), allEdges[i].getSecond(), allEdges[i].label); nb_removed++; } fclose(output); fclose(removed_out); fprintf(stdout, "%d out of %d edges removed\n", nb_removed, (int)allEdges.size()); return 0; } kissplice-2.6.2/modules/LabelledCEdge.cpp000066400000000000000000000127651423273700400202740ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ // =========================================================================== // Include Libraries // =========================================================================== #include // =========================================================================== // Include Project Files // =========================================================================== #include "LabelledCEdge.h" // =========================================================================== // Declare Used Namespaces // =========================================================================== //############################################################################ // # // Class LabelledCEdge # // # //############################################################################ // =========================================================================== // Static attributes // =========================================================================== // =========================================================================== // Constructors // =========================================================================== LabelledCEdge::LabelledCEdge( const LabelledCEdge& model ) : CEdge( model ) { strcpy( label, model.label ); } /*! * */ LabelledCEdge::LabelledCEdge( int f, int s, char* _label ) : CEdge( f, s ) { strcpy( label, _label ); }; LabelledCEdge::LabelledCEdge( CEdge& edge, char* _label ) : CEdge( edge ) { strcpy( label, _label ); }; // =========================================================================== // Destructor // =========================================================================== // =========================================================================== // Operators // =========================================================================== bool LabelledCEdge::operator<( const LabelledCEdge& that ) const { if ( static_cast( *this ) == static_cast( that ) ) { if ( label[0] == that.label[0] ) { return label[1] < that.label[1]; } return label[0] < that.label[0]; } return static_cast( *this ) < static_cast( that ); } bool LabelledCEdge::operator==( const LabelledCEdge& that ) const { return static_cast( *this ) == static_cast( that ) && label[0] == that.label[0] && label[1] == that.label[1]; } // =========================================================================== // Public Methods // =========================================================================== // =========================================================================== // Protected Methods // =========================================================================== // =========================================================================== // Non inline accessors // =========================================================================== kissplice-2.6.2/modules/LabelledCEdge.h000066400000000000000000000157621423273700400177410ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #ifndef LABELLED_EDGE_H #define LABELLED_EDGE_H // =========================================================================== // Include Libraries // =========================================================================== #include #include // =========================================================================== // Include Project Files // =========================================================================== #include "CEdge.h" // =========================================================================== // Class declarations // =========================================================================== // =========================================================================== // Declare Used Namespaces // =========================================================================== //! Labelled edge /*! * \brief Class meant to be used by NGraph to represent edges. * Inherits from CEdge class. Contains: * a pair of int (u -> v edge), each int (u or v) being a node. * a label representing, most of the time, the direction of the edge (FR, RR, RF, FF) */ class LabelledCEdge : public CEdge { public : // ======================================================================= // Enums // ======================================================================= // ======================================================================= // Constructors // ======================================================================= LabelledCEdge( const LabelledCEdge& model ); LabelledCEdge( int f, int s, char* _label ); LabelledCEdge( CEdge& edge, char* _label ); // ======================================================================= // Destructor // ======================================================================= // ======================================================================= // Accessors: getters // ======================================================================= // ======================================================================= // Accessors: setters // ======================================================================= // ======================================================================= // Operators // ======================================================================= bool operator<( const LabelledCEdge& that ) const; bool operator==( const LabelledCEdge& that ) const; // ======================================================================= // Public Methods // ======================================================================= // ======================================================================= // Public Attributes // ======================================================================= char label[3]; protected : // ======================================================================= // Forbidden Constructors // ======================================================================= LabelledCEdge( void ) { printf( "%s:%d: error: call to forbidden constructor.\n", __FILE__, __LINE__ ); exit( EXIT_FAILURE ); }; // ======================================================================= // Protected Methods // ======================================================================= // ======================================================================= // Protected Attributes // ======================================================================= }; // =========================================================================== // Getters' definitions // =========================================================================== // =========================================================================== // Setters' definitions // =========================================================================== // =========================================================================== // Inline Operators' definitions // =========================================================================== // =========================================================================== // Inline functions' definition // =========================================================================== #endif // LABELLED_EDGE_H kissplice-2.6.2/modules/NEdge.h000066400000000000000000000132211423273700400163130ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ #ifndef NEDGE_H #define NEDGE_H // =========================================================================== // Include Libraries // =========================================================================== #include #include #include // =========================================================================== // Include Project Files // =========================================================================== #include "Utils.h" /* The node class is used into the adjacency list of the NGraph object. It keeps the whole information of the link between a node u and its outgoing neighbours: the index of the node v and the label of the arc linking u to v. */ class NEdge { public : // ======================================================================= // Enums // ======================================================================= // ======================================================================= // Constructors // ======================================================================= NEdge() = delete; NEdge( const NEdge& ) = default; NEdge( int node, string labels ) : m_node(node), m_labels(std::move(labels)) {} NEdge( int node, char label1, char label2 ) : m_node(node) { char buf[3] = { label1, label2, '\0' }; m_labels = std::string( buf ); } // ======================================================================= // Accessors: getters // ======================================================================= inline int get_node( void ) const { return m_node; } inline string get_labels( void ) const { return m_labels; } // ======================================================================= // Accessors: setters // ======================================================================= inline void set_node( int node ) { m_node = node; } inline void set_labels( string labels ) { m_labels = std::move(labels); } // ======================================================================= // Operators // ======================================================================= // ======================================================================= // Public Methods // ======================================================================= void add_label( const string& label ) { m_labels += label; } void del_all_but_first() { m_labels.erase( 2 ); } void revert_dir( int i ) { m_labels[i] = reverse_dir(m_labels[i]); } // ======================================================================= // Public Attributes // ======================================================================= protected : // ======================================================================= // Protected Methods // ======================================================================= // ======================================================================= // Protected Attributes // ======================================================================= //! Index of the node int m_node; //! Label of the incoming edge string m_labels; }; #endif // NEDGE_H kissplice-2.6.2/modules/NGraph.cpp000066400000000000000000001054311423273700400170500ustar00rootroot00000000000000/* *************************************************************************** * * KisSplice * de-novo calling alternative splicing events from RNA-seq data. * * *************************************************************************** * * Copyright INRIA * contributors : Vincent Lacroix * Pierre Peterlongo * Gustavo Sacomoto * Vincent Miele * Alice Julien-Laferriere * David Parsons * * pierre.peterlongo@inria.fr * vincent.lacroix@univ-lyon1.fr * * This software is a computer program whose purpose is to detect alternative * splicing events from RNA-seq data. * * This software is governed by the CeCILL license under French law and * abiding by the rules of distribution of free software. You can use, * modify and/ or redistribute the software under the terms of the CeCILL * license as circulated by CEA, CNRS and INRIA at the following URL * "http://www.cecill.info". * As a counterpart to the access to the source code and rights to copy, * modify and redistribute granted by the license, users are provided only * with a limited warranty and the software's author, the holder of the * economic rights, and the successive licensors have only limited * liability. * In this respect, the user's attention is drawn to the risks associated * with loading, using, modifying and/or developing or reproducing the * software by the user in light of its specific status of free software, * that may mean that it is complicated to manipulate, and that also * therefore means that it is reserved for developers and experienced * professionals having in-depth computer knowledge. Users are therefore * encouraged to load and test the software's suitability as regards their * requirements in conditions enabling the security of their systems and/or * data to be ensured and, more generally, to use and operate it in the * same conditions as regards security. * * The fact that you are presently reading this means that you have had * knowledge of the CeCILL license and that you accept its terms. */ // =========================================================================== // Include Libraries // =========================================================================== #include #include #include #include #include #include #include #include #include #include #include // =========================================================================== // Include Project Files // =========================================================================== #include "NGraph.h" #include "debug.h" #include "CycleCompression.h" #include "Utils.h" // =========================================================================== // Declare Used Namespaces // =========================================================================== using namespace std; // =========================================================================== // Constant // =========================================================================== #define MAX 1024 //############################################################################ // # // Class NGraph # // # //############################################################################ // =========================================================================== // Constructors // =========================================================================== NGraph::NGraph( int kValue, int outputtedSnps ) { _kValue = kValue; _nbOutput = outputtedSnps; } /*! TODO : check : Create a NGraph for the BCC passed in edges ? all_edges ? */ NGraph::NGraph( CGraph& cgraph, vector& seqs, vector& all_edges, vector& edges ) { _nbOutput = 0; for ( int i = 0 ; i < (int)edges.size() ; i++ ) { int u = edges[i].getFirst(); int v = edges[i].getSecond(); //~ printf( "contructing %s %s (0x%x, 0x%x)\n", seqs[u], seqs[v], seqs[u], seqs[v] ); //~ getchar(); insert_node( to_str(u), to_str( seqs[u] ) ); insert_node( to_str(v), to_str( seqs[v] ) ); insert_bidirected_edges( all_edges, edges[i] ); edges[i].swap_ends(); insert_bidirected_edges( all_edges, edges[i] ); } _kValue = cgraph.k_value; expand_parallel_edges(); } // =========================================================================== // Public Methods // =========================================================================== void NGraph::insert_empty_node( string u ) { _strToNode[u] = _nodeToStr.size(); // _nodeToStr.size() corresponds to the current number of nodes in the graph _nodeToStr.push_back(u); _nodes.push_back( NNode() ); } // Insert the new edge u->v (label). // If the edge u->v is already present with a different label, concatenate