From 0507ed5e0bb82fb4cfb42ff6ff03664461643177 Mon Sep 17 00:00:00 2001 From: "christoph.stritt@unibas.ch" <christoph.stritt@unibas.ch> Date: Wed, 18 Oct 2023 14:49:47 +0200 Subject: [PATCH] Parameter outdir added to config --- .gitignore | 3 +- README.md | 34 +- assembly/README.md | 36 +- assembly/config/config.yaml | 10 +- assembly/config/samples.tsv | 0 assembly/container/README.md | 6 +- assembly/container/assemblySC.def | 0 assembly/container/conda-linux-64.lock | 161 +- assembly/container/conda-lock.yml | 3859 ++++++++--------- assembly/container/environment.yml | 2 +- assembly/resources/H37Rv.gbf | 0 assembly/workflow/Snakefile | 27 +- assembly/workflow/rules/annotate.smk | 47 +- assembly/workflow/rules/assemble.smk | 9 +- assembly/workflow/rules/circularize.smk | 44 +- assembly/workflow/rules/cleanup.smk | 7 - assembly/workflow/rules/common.smk | 0 assembly/workflow/rules/mapreads.smk | 0 assembly/workflow/rules/readQC.smk | 6 +- assembly/workflow/rules/summarize.smk | 15 +- fig.png | Bin 42853 -> 0 bytes variantcalling/README.md | 8 + variantcalling/config/biopython.yaml | 0 variantcalling/config/config.yaml | 0 variantcalling/config/samples.txt | 0 variantcalling/workflow/Snakefile | 0 .../workflow/scripts/combine_assemblies.py | 0 27 files changed, 1948 insertions(+), 2326 deletions(-) mode change 100644 => 100755 .gitignore mode change 100644 => 100755 README.md mode change 100644 => 100755 assembly/README.md mode change 100644 => 100755 assembly/config/config.yaml mode change 100644 => 100755 assembly/config/samples.tsv mode change 100644 => 100755 assembly/container/README.md mode change 100644 => 100755 assembly/container/assemblySC.def mode change 100644 => 100755 assembly/container/conda-linux-64.lock mode change 100644 => 100755 assembly/container/conda-lock.yml mode change 100644 => 100755 assembly/container/environment.yml mode change 100644 => 100755 assembly/resources/H37Rv.gbf mode change 100644 => 100755 assembly/workflow/rules/annotate.smk mode change 100644 => 100755 assembly/workflow/rules/assemble.smk mode change 100644 => 100755 assembly/workflow/rules/circularize.smk delete mode 100644 assembly/workflow/rules/cleanup.smk mode change 100644 => 100755 assembly/workflow/rules/common.smk mode change 100644 => 100755 assembly/workflow/rules/mapreads.smk mode change 100644 => 100755 assembly/workflow/rules/readQC.smk mode change 100644 => 100755 assembly/workflow/rules/summarize.smk delete mode 100644 fig.png mode change 100644 => 100755 variantcalling/README.md mode change 100644 => 100755 variantcalling/config/biopython.yaml mode change 100644 => 100755 variantcalling/config/config.yaml mode change 100644 => 100755 variantcalling/config/samples.txt mode change 100644 => 100755 variantcalling/workflow/Snakefile mode change 100644 => 100755 variantcalling/workflow/scripts/combine_assemblies.py diff --git a/.gitignore b/.gitignore old mode 100644 new mode 100755 index 2defeaa..d8d6f29 --- a/.gitignore +++ b/.gitignore @@ -1,4 +1,5 @@ pangenome assembly/container/assemblySC.sif assembly/.snakemake -assembly/rulegraph.pdf \ No newline at end of file +assembly/rulegraph.pdf +facienda.md \ No newline at end of file diff --git a/README.md b/README.md old mode 100644 new mode 100755 index a1293f5..1efd40a --- a/README.md +++ b/README.md @@ -6,15 +6,39 @@ This folder contains two Snakemake workflows: This is ongoing work, some things will change. + ## Requirements -Running the workflows requires [Snakemake](https://snakemake.readthedocs.io) and [Singularity](https://docs.sylabs.io/guides/latest/user-guide/). Both are installed on sciCORE and can be loaded as modules: +On the sciCORE cluster, the pipeline is installed in the GROUP folder (**/scicore/home/gagneux/GROUP/PacbioSnake**) and ready to run. + +In other contexts, four things need to be set up before the pipeline can be run: + + 1. Install Snakemake and Singularity + 2. Build the singularity container for the assembly pipeline + 3. Download the bakta database for genome annotation + 4. Pull the singularity container for the variant calling pipeline + +These steps are detailed below. + + +### 1. Install Snakemake and Singularity +As described on the [Snakemake](https://snakemake.readthedocs.io) and the [Singularity](https://docs.sylabs.io/guides/latest/user-guide/) sites. + +### 2. Build the singularity container for the assembly pipeline ``` -ml snakemake/6.6.1-foss-2021a -ml Singularity/2.6.1 +cd assembly/container +sudo singularity build assemblySC.sif assemblySC.def ``` -<img src="fig.png" width="50%" height="50%"> + +### 3. Download the bakta database for genome annotation +Light-weight (1.3 Gb) and full (33.1 Gb) databases for the *bakta* annotation tool can be downloaded from https://zenodo.org/records/7669534. +The extracted folder should be located at assembly/resources/bakta_db/. Otherwise the path to the database can be modified in the assembly config file. + + +### 4. Pull the singularity container for the variant calling pipeline +``` +singularity pull docker://ghcr.io/pangenome/pggb:latest +``` -*Section of a genome graph constructed from diverse MTBC genomes. It shows a nested structural variant: an IS6110 insertion within the TBD1 region.* diff --git a/assembly/README.md b/assembly/README.md old mode 100644 new mode 100755 index e55f57e..f3b9227 --- a/assembly/README.md +++ b/assembly/README.md @@ -6,16 +6,6 @@ The user needs to modify two things to run the workflow on her samples: - the file config/config.yaml, which contains global options for the analysis -## container -Problem: 1.6 gb container on gitlab? Building or loading container requires sudo... - - -``` -sudo singularity build assemblySMK.sif assemblySMK.def -``` - - - ## samples.tsv This is a tab-separated table with a header and two colums (see example): the first containing the sample names, which will be used to name the assemblies; the second with the absolute paths to the fastq files. @@ -57,20 +47,30 @@ Important: singularity containers most be given access to the file locations thr It's convenient to run snakemake in a screen, so we can do other things on scicore while it's running and occasionally check the progress. -Important snakemake arguments: - --jobs: - --cluster ``` +# Open screen screen -r assembly +# Load Snakemake module +ml snakemake/6.6.1-foss-2021a + +# Dry run + + +# Real run snakemake \ + --config samples=1.5 \ + --config outdir=./results \ + --jobs 4 - -k \ - --latency-wait 60 \ --use-singularity --singularity-args "--bind /scicore/home/gagneux/GROUP/tbresearch/genomes/IN_PROGRESS/PacBio_genomes/Gagneux --bind /scicore/home/gagneux/stritt0001/TB/projects/pacbio_microscale/ --bind /scratch" \ - --cluster "sbatch --job-name=hifi_simul --cpus-per-task=4 --mem-per-cpu=4G --time=06:00:00 --qos=6hours --output=hifi_simul.o%j --error=hifi_simul.e%j" + --cluster "sbatch --job-name=pbassembly --cpus-per-task=4 --mem-per-cpu=4G --time=06:00:00 --qos=6hours --output=pbassembly.o%j --error=pbassembly.e%j" -``` -Leave the screen with CTRL+a+d. \ No newline at end of file +# Leave the screen: CTRL+a+d + +# Return to the screen +screen -r assembly + +``` \ No newline at end of file diff --git a/assembly/config/config.yaml b/assembly/config/config.yaml old mode 100644 new mode 100755 index 6b82edb..e3405c4 --- a/assembly/config/config.yaml +++ b/assembly/config/config.yaml @@ -2,16 +2,16 @@ # ############################## -project_directory: ./results -output_prefix: pb_bernese samples: config/samples.tsv +outdir: ./results +output_prefix: pb_bernese ref: genome_size: 4.4m gbf: resources/H37Rv.gbf -threads: 4 +bakta_db: resources/bakta_db -keep_intermediate: "Yes" +threads_per_job: 4 -mapreads: "No" +keep_intermediate: "Yes" diff --git a/assembly/config/samples.tsv b/assembly/config/samples.tsv old mode 100644 new mode 100755 diff --git a/assembly/container/README.md b/assembly/container/README.md old mode 100644 new mode 100755 index 83dbaae..d275f24 --- a/assembly/container/README.md +++ b/assembly/container/README.md @@ -34,8 +34,4 @@ conda env update --file environment.yml --prune conda-lock --mamba -f environment.yml -p linux-64 conda-lock render -p linux-64 # allows using mamba create --file conda-linux-64.lock -Problem: 1.6 gb container on gitlab? Building or loading container requires sudo... - -``` -sudo singularity build assemblySC.sif assemblySC.def -``` \ No newline at end of file +# The conda-linux-64.lock file is then used in assemblySC.def to build the container diff --git a/assembly/container/assemblySC.def b/assembly/container/assemblySC.def old mode 100644 new mode 100755 diff --git a/assembly/container/conda-linux-64.lock b/assembly/container/conda-linux-64.lock old mode 100644 new mode 100755 index 7a872e3..15afcb4 --- a/assembly/container/conda-linux-64.lock +++ b/assembly/container/conda-linux-64.lock @@ -1,14 +1,12 @@ # Generated by conda-lock. # platform: linux-64 -# input_hash: 926ab17aedcc8b15a4be8db176a2c48cafc2fcb2c9f778737a63eb301e4e346e +# input_hash: 2e06febb1f32f5d3e52edc793538b8ec57542223dda338ed8fb24ab1ce919365 @EXPLICIT https://conda.anaconda.org/conda-forge/linux-64/_libgcc_mutex-0.1-conda_forge.tar.bz2#d7c89558ba9fa0495403155b64376d81 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https://repo.anaconda.com/pkgs/main/noarch/tzdata-2023c-h04d1e81_0.conda#29db02adf8808f7c64642cead3e28acd @@ -19,21 +17,18 @@ https://conda.anaconda.org/conda-forge/linux-64/_openmp_mutex-4.5-2_gnu.tar.bz2# https://repo.anaconda.com/pkgs/main/noarch/sysroot_linux-64-2.17-h57e8cba_10.tar.bz2#daa11b19c141284d62536ba03f4c73aa https://conda.anaconda.org/conda-forge/linux-64/libgcc-ng-13.2.0-h807b86a_2.conda#c28003b0be0494f9a7664389146716ff https://conda.anaconda.org/bioconda/linux-64/aragorn-1.2.41-h031d066_1.tar.bz2#d4ed7d256f07678d839cadedd1d49443 -https://conda.anaconda.org/conda-forge/linux-64/argtable2-2.13-h14c3975_1001.tar.bz2#b9e0ac5d747ceab0c8b85a470358e1a6 https://repo.anaconda.com/pkgs/main/linux-64/bzip2-1.0.8-h7b6447c_0.conda#9303f4af7c004e069bae22bde8d800ee https://repo.anaconda.com/pkgs/main/linux-64/c-ares-1.19.1-h5eee18b_0.conda#6cfbce52273a1cb888821f18ceaa83c4 -https://conda.anaconda.org/bioconda/linux-64/clustalw-2.1-h4ac6f70_9.tar.bz2#919194f6f7d733afff4d3c6f54e08d07 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https://repo.anaconda.com/pkgs/main/linux-64/libev-4.33-h7f8727e_1.conda#5065620db4393fb549f30114a33897d1 https://repo.anaconda.com/pkgs/main/linux-64/libffi-3.4.4-h6a678d5_0.conda#06e288f9250abef59b9a367d151fc339 -https://repo.anaconda.com/pkgs/main/linux-64/libidn11-1.33-h7b6447c_0.conda#75d5c232628eb6701d7a491716243a1f https://repo.anaconda.com/pkgs/main/linux-64/libnsl-2.0.0-h5eee18b_0.conda#55e3f1b0c2a70454cdf1a5d64fb44752 https://repo.anaconda.com/pkgs/main/linux-64/libopenblas-0.3.21-h043d6bf_0.conda#7f7324dcc3c4761a14f3e4ac443235a7 https://repo.anaconda.com/pkgs/main/linux-64/libunistring-0.9.10-h27cfd23_0.conda#5b6ed6cd1b1ca3a383daef1d6464d02a @@ -43,18 +38,14 @@ https://repo.anaconda.com/pkgs/main/linux-64/libxcb-1.15-h7f8727e_0.conda#ada518 https://conda.anaconda.org/conda-forge/linux-64/libzlib-1.2.13-hd590300_5.conda#f36c115f1ee199da648e0597ec2047ad https://conda.anaconda.org/conda-forge/linux-64/llvm-openmp-8.0.1-hc9558a2_0.tar.bz2#67590caab043d6d7ffc371f9cced7848 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DO NOT EDIT! +# This lock file was generated by conda-lock (https://github.com/conda/conda-lock). DO NOT EDIT! # # A "lock file" contains a concrete list of package versions (with checksums) to be installed. Unlike # e.g. `conda env create`, the resulting environment will not change as new package versions become @@ -9,8 +9,11 @@ # To update a single package to the latest version compatible with the version constraints in the source: # conda-lock lock --lockfile conda-lock.yml --update PACKAGE # To re-solve the entire environment, e.g. after changing a version constraint in the source file: -# conda-lock -f /home/cristobal/TB/projects/pacbio_microscale/gitlab/PacbioSnake/assembly/container/environment.yml --lockfile conda-lock.yml +# conda-lock -f /home/cristobal/TB/projects/pacbio_microscale/gitlab/PacbioSnake/assembly/container/environment.yml -f environment.yml --lockfile conda-lock.yml +version: 1 metadata: + content_hash: + linux-64: 2e06febb1f32f5d3e52edc793538b8ec57542223dda338ed8fb24ab1ce919365 channels: - url: defaults used_env_vars: [] @@ -20,1778 +23,1513 @@ metadata: used_env_vars: [] - url: r used_env_vars: [] - content_hash: - linux-64: 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optional: false +- name: matplotlib + version: 3.7.2 + manager: conda platform: linux-64 - url: https://repo.anaconda.com/pkgs/main/linux-64/pyqt-5.15.7-py310h6a678d5_1.conda - version: 5.15.7 -- category: main dependencies: matplotlib-base: '>=3.7.2,<3.7.3.0a0' pyqt: '>=5' python: '>=3.10,<3.11.0a0' tornado: '>=5' + url: https://repo.anaconda.com/pkgs/main/linux-64/matplotlib-3.7.2-py310h06a4308_0.conda hash: md5: d82ab76c17c1328f1966a198e72ee5e7 sha256: e3b148b85eb0214595c236791ae1bf24f4d058151a079703b10316a17b434f7c - manager: conda - name: matplotlib + category: main optional: false +- name: ncbi-vdb + version: 3.0.8 + manager: conda platform: linux-64 - url: https://repo.anaconda.com/pkgs/main/linux-64/matplotlib-3.7.2-py310h06a4308_0.conda - version: 3.7.2 -- category: main dependencies: ca-certificates: '' curl: '' @@ -3766,45 +3484,32 @@ package: perl: '' perl-uri: '' perl-xml-libxml: '' + url: https://conda.anaconda.org/bioconda/linux-64/ncbi-vdb-3.0.8-hdbdd923_0.tar.bz2 hash: md5: 3b58cf2b702f96d812c67466456fe251 sha256: 2a16ded5977c91b379fcf7ec4cc7d033d5a0b66669ab438579c92f10e99ec186 - manager: conda - name: ncbi-vdb + category: main optional: false +- name: requests + version: 2.31.0 + manager: conda platform: linux-64 - url: https://conda.anaconda.org/bioconda/linux-64/ncbi-vdb-3.0.8-hdbdd923_0.tar.bz2 - version: 3.0.8 -- category: main dependencies: - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-base: '' - perl-digest-md5: '' - perl-encode: '' - perl-encode-locale: '' - perl-file-listing: '' - perl-html-parser: '' - perl-http-cookies: '' - perl-http-daemon: '' - perl-http-date: '' - perl-http-negotiate: '' - perl-lwp-mediatypes: '' - perl-mime-base64: '' - perl-net-http: '>=6.18' - perl-ntlm: '' - perl-try-tiny: '' - perl-uri: '' - perl-www-robotrules: '' + certifi: '>=2017.4.17' + charset-normalizer: '>=2,<4' + idna: '>=2.5,<4' + python: '>=3.10,<3.11.0a0' + urllib3: '>=1.21.1,<2' + url: https://repo.anaconda.com/pkgs/main/linux-64/requests-2.31.0-py310h06a4308_0.conda hash: - md5: 9493077e9e1dae0017f0d4cd9baf7de6 - sha256: cb997a7d5ff37db0dafbcf7650645e64d23bfe0f1109c34f54e064f3bfe76a2c - manager: conda - name: perl-libwww-perl + md5: 81e8293939370975e4887b12336e1b89 + sha256: 23417b09593a32fa446b36fbf5949ae6a6aa59e4b66091850f973fea92c6992b + category: main optional: false +- name: blast + version: 2.14.1 + manager: conda platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-libwww-perl-6.67-pl5321hdfd78af_0.tar.bz2 - version: '6.67' -- category: main dependencies: bzip2: '>=1.0.8,<2.0a0' curl: '' @@ -3819,141 +3524,69 @@ package: perl-json: '' perl-list-moreutils: '' zlib: '' + url: https://conda.anaconda.org/bioconda/linux-64/blast-2.14.1-pl5321h6f7f691_0.tar.bz2 hash: md5: 717b2ae9b971e769e00b984e37fe8947 sha256: a83d7e119a32a39d9d839c691d811dcea091e06dd4eac3032ab3244be606baa6 - manager: conda - name: blast - optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/linux-64/blast-2.14.1-pl5321h6f7f691_0.tar.bz2 - version: 2.14.1 -- category: main - dependencies: - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-http-message: '>=6.18' - perl-libwww-perl: '' - perl-libxml-perl: '' - perl-xml-parser: '' - perl-xml-regexp: '' - hash: - md5: 45b7ce435727456727f45b3e2121b23b - sha256: 5d51356aa3d2f46ff6bf060567bbaa30b4a9dcbab8d5abf9cbaa50c8a10aa369 - manager: conda - name: perl-xml-dom + category: main optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-xml-dom-1.46-pl5321hdfd78af_1.tar.bz2 - version: '1.46' -- category: main - dependencies: - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-xml-dom: '' - perl-xml-xpathengine: '' - hash: - md5: b959a7e6e307abaec3f06b0bc007809a - sha256: d50d8bf14f9562cefe230f2333b26475ffac6b859d72bae19afb02ca52a0245b +- name: diamond + version: 2.1.8 manager: conda - name: perl-xml-dom-xpath - optional: false platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-xml-dom-xpath-0.14-pl5321hdfd78af_2.tar.bz2 - version: '0.14' -- category: main dependencies: blast: '' - clustalo: '' - clustalw: '' - libgcc-ng: '>=4.9' - libgfortran: '>=3.0' - libstdcxx-ng: '>=4.9' - mafft: '' - muscle: '' - poa: '' - viennarna: '' + libgcc-ng: '>=12' + libstdcxx-ng: '>=12' + libzlib: '>=1.2.13,<1.3.0a0' + zlib: '' + url: https://conda.anaconda.org/bioconda/linux-64/diamond-2.1.8-h43eeafb_0.tar.bz2 hash: - md5: a0e9a4fa83c8c2cb485f248cf5ec0a34 - sha256: 33055cffed0ad42ee41306759f9b10c8e0a80f44850b7a7b8a67c4e034ef2839 - manager: conda - name: t-coffee + md5: 86ccdf1d01687ea915c5a805ab6aefbc + sha256: 341c337124b3930df384526f676956f42f866bc9669f0ea912c1fea88b5986ae + category: main optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/linux-64/t-coffee-12.00.7fb08c2-h26a2512_0.tar.bz2 - version: 12.00.7fb08c2 -- category: main - dependencies: - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-bioperl-core: '' - perl-tree-dag_node: '' - perl-uri: '' - perl-xml-dom: '' - perl-xml-dom-xpath: '' - hash: - md5: 1b4b260b45e9cac33fcb45918cda428b - sha256: d63661b5a81ddc9c80dba2992fa9391e2ca9e4da15c201113dee83becbc6a4ef +- name: ncbi-amrfinderplus + version: 3.11.26 manager: conda - name: perl-bio-featureio - optional: false platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-bio-featureio-1.6.905-pl5321hdfd78af_4.tar.bz2 - version: 1.6.905 -- category: main dependencies: - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-bioperl-run: '' - t-coffee: '' - hash: - md5: 932416c9ba6a9af43609e04ec5952574 - sha256: 0ffaced1dbe89e5bd626e2d467822848b66e084d4b3fa6a21ba9af26a7133aaa - manager: conda - name: perl-bio-tools-run-alignment-tcoffee + blast: '>=2.9' + curl: '' + hmmer: '>=3.2' + libcurl: '>=8.4.0,<9.0a0' + libgcc-ng: '>=12' + libstdcxx-ng: '>=12' + url: https://conda.anaconda.org/bioconda/linux-64/ncbi-amrfinderplus-3.11.26-h283d18e_0.tar.bz2 + hash: + md5: 531aa783d5e124a7168a0107a8ba6bd9 + sha256: 7c88402f254d3ede232c9b51803d3bee06c9809d31c746e51c94ea6ecb8409bf + category: main + optional: false +- name: bakta + version: 1.8.2 + manager: conda + platform: linux-64 + dependencies: + alive-progress: 3.0.1 + aragorn: '>=1.2.41' + biopython: '>=1.78' + blast: '>=2.12.0' + circos: '>=0.69.8' + diamond: '>=2.0.14' + infernal: '>=1.1.4' + ncbi-amrfinderplus: '>=3.11.2' + piler-cr: '' + pyhmmer: '>=0.10.0' + pyrodigal: '>=2.1.0,<3.0' + python: '>=3.8,<3.11' + pyyaml: '>=6.0' + requests: '>=2.25.1' + trnascan-se: '>=2.0.11' + xopen: '>=1.5.0' + url: https://conda.anaconda.org/bioconda/noarch/bakta-1.8.2-pyhdfd78af_1.tar.bz2 + hash: + md5: fcb919368132b69a0bba9157d4fa16a0 + sha256: 344e063daf3c3cd21d289e85f7958e5cbdb726d63742076dff7401d9bbcfc07f + category: main optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-bio-tools-run-alignment-tcoffee-1.7.4-pl5321hdfd78af_5.tar.bz2 - version: 1.7.4 -- category: main - dependencies: - perl: '' - perl-bio-asn1-entrezgene: '' - perl-bio-coordinate: '' - perl-bio-featureio: '' - perl-bio-samtools: '' - perl-bio-searchio-hmmer: '' - perl-bio-tools-phylo-paml: '' - perl-bio-tools-run-alignment-clustalw: '' - perl-bio-tools-run-alignment-tcoffee: '' - perl-bioperl-core: 1.7.8.* - perl-bioperl-run: '' - hash: - md5: a693939710e572d0f85f6058b534e319 - sha256: 9632f59b230f0b5c9ba168ec742ba4dabc77b2cdac63ac71aaa8846f26bd4cb7 - manager: conda - name: perl-bioperl - optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/perl-bioperl-1.7.8-hdfd78af_1.tar.bz2 - version: 1.7.8 -- category: main - dependencies: - aragorn: '>=1.2' - barrnap: '>=0.7' - blast: '>=2.7.1' - hmmer: '>=3.1b2' - infernal: '>=1.1.2' - minced: '>=0.3' - parallel: '>=20180522' - perl: '>=5.32.1,<6.0a0 *_perl5' - perl-bioperl: '>=1.7.2' - perl-xml-simple: '' - prodigal: '>=2.6' - tbl2asn-forever: '>=25.7' - hash: - md5: 6aea788d2ca74ab50d6816e7e0a75da2 - sha256: 654e7ab399907d0153052dc2f729d4b8db5de6c819686d2dbb8a4890782595c4 - manager: conda - name: prokka - optional: false - platform: linux-64 - url: https://conda.anaconda.org/bioconda/noarch/prokka-1.14.6-pl5321hdfd78af_5.tar.bz2 - version: 1.14.6 -version: 1 diff --git a/assembly/container/environment.yml b/assembly/container/environment.yml old mode 100644 new mode 100755 index cbebc99..adfc0d2 --- a/assembly/container/environment.yml +++ b/assembly/container/environment.yml @@ -9,7 +9,7 @@ dependencies: - samtools - minimap2 - circlator - - prokka + - bakta - numpy - scipy - matplotlib diff --git a/assembly/resources/H37Rv.gbf b/assembly/resources/H37Rv.gbf old mode 100644 new mode 100755 diff --git a/assembly/workflow/Snakefile b/assembly/workflow/Snakefile index b18e49c..aa6153d 100755 --- a/assembly/workflow/Snakefile +++ b/assembly/workflow/Snakefile @@ -13,22 +13,11 @@ include: "rules/annotate.smk" include: "rules/summarize.smk" -if config["mapreads"] == "yes": - - include: "rules/mapreads.smk" - - rule all: - input: - "results/read_summaries.tsv", - "results/assembly_summaries.tsv", - expand("results/{sample}/prokka/{sample}.gff", sample = samples.keys()), - expand("results/{sample}/remapping/shortreads.bam", sample = samples.keys()), - expand("results/{sample}/remapping/longreads.bam", sample = samples.keys()) - -else: - rule all: - input: - "results/read_summaries.tsv", - "results/assembly_summaries.tsv", - expand("results/{sample}/prokka/{sample}.gff", sample = samples.keys()) - +include: "rules/mapreads.smk" + +rule all: + input: + config["outdir"] + "/read_summaries.tsv", + config["outdir"] + "/assembly_summaries.tsv", + expand(config["outdir"] + "/{sample}/{sample}.gff", sample = samples.keys()), + expand(config["outdir"] + "/{sample}/remapping/longreads.bam", sample = samples.keys()) diff --git a/assembly/workflow/rules/annotate.smk b/assembly/workflow/rules/annotate.smk old mode 100644 new mode 100755 index 6a54e45..b8a55bf --- a/assembly/workflow/rules/annotate.smk +++ b/assembly/workflow/rules/annotate.smk @@ -1,27 +1,26 @@ -rule prokka: - input: "results/{sample}/assembly.circularized.renamed.fasta" - output: "results/{sample}/prokka/{sample}.gff" - params: - refgbf = config["ref"]["gbf"], - asm = "{sample}", - outdir = "results/{sample}/prokka" - threads: config["threads"] - envmodules: "prokka/1.14.5-gompi-2021a" - shell: - """ - prokka {input} \ - --force \ - --kingdom Bacteria \ - --genus Mycobacterium \ - --species tuberculosis \ - --strain {params.asm} \ - --addgenes \ - --proteins {params.refgbf} \ - --prefix {params.asm} \ - --locustag {params.asm} \ - --cpus {threads} \ - --outdir {params.outdir} +rule bakta: + input: config["outdir"] + "/{sample}/assembly.circularized.renamed.fasta" + output: config["outdir"] + "/{sample}/{sample}.gff" + params: + bakta_db = config["bakta_db"], + refgbf = config["ref"]["gbf"], + asm = "{sample}", + outdir = config["outdir"] + "/{sample}/", + threads = config["threads_per_job"] - """ + shell: + """ + bakta {input} \ + --db {params.bakta_db} \ + --prefix {params.asm} \ + --genus Mycobacterium \ + --species tuberculosis \ + --strain {params.sample} \ + --locus-tag {params.sample} \ + --complete --compatible \ + --proteins {params.refgbf} \ + --keep-contig-headers \ + --threads {params.threads} + """ diff --git a/assembly/workflow/rules/assemble.smk b/assembly/workflow/rules/assemble.smk old mode 100644 new mode 100755 index cec994b..90d09ff --- a/assembly/workflow/rules/assemble.smk +++ b/assembly/workflow/rules/assemble.smk @@ -1,11 +1,13 @@ rule flye: input: lambda wildcards: samples[wildcards.sample].longread_fastq - output: "results/{sample}/assembly.fasta", "results/{sample}/assembly_info.txt" - threads: config["threads"] + output: + config["outdir"] + "/{sample}/flye/assembly.fasta", + config["outdir"] + "/{sample}/flye/assembly_info.txt" + threads: config["threads_per_job"] params: genome_size = config["ref"]["genome_size"], - outdir = "results/{sample}" + outdir = config["outdir"] + "/{sample}/flye" shell: """ @@ -15,4 +17,3 @@ rule flye: --threads {threads} \ --out-dir {params.outdir} """ - diff --git a/assembly/workflow/rules/circularize.smk b/assembly/workflow/rules/circularize.smk old mode 100644 new mode 100755 index cce5cb3..befa762 --- a/assembly/workflow/rules/circularize.smk +++ b/assembly/workflow/rules/circularize.smk @@ -1,11 +1,11 @@ rule circlator_mapreads: input: - assembly = 'results/{sample}/assembly.fasta', + assembly = config["outdir"] + "/{sample}/assembly.fasta", reads = lambda wildcards: expand(samples[wildcards.sample].longread_fastq) - output: "results/{sample}/circlator/01.mapreads.bam" + output: config["outdir"] + "/{sample}/circlator/01.mapreads.bam" params: - threads = config["threads"] + threads = config["threads_per_job"] shell: """ @@ -14,10 +14,10 @@ rule circlator_mapreads: """ rule circlator_bam2reads: - input: "results/{sample}/circlator/01.mapreads.bam" - output: "results/{sample}/circlator/02.bam2reads.fasta" + input: config["outdir"] + "/{sample}/circlator/01.mapreads.bam" + output: config["outdir"] +"/{sample}/circlator/02.bam2reads.fasta" params: - output_pref = "results/{sample}/circlator/02.bam2reads" + output_pref = config["outdir"]+ "/{sample}/circlator/02.bam2reads" shell: """ circlator bam2reads {input} {params.output_pref} --discard_unmapped @@ -25,10 +25,10 @@ rule circlator_bam2reads: """ rule circlator_localassembly: - input: "results/{sample}/circlator/02.bam2reads.fasta" - output: "results/{sample}/circlator/03.assemble/assembly.fasta" + input: config["outdir"] + "/{sample}/circlator/02.bam2reads.fasta" + output: config["outdir"] + "/{sample}/circlator/03.assemble/assembly.fasta" params: - outdir = "results/{sample}/circlator/03.assemble", + outdir = config["outdir"] + "/{sample}/circlator/03.assemble", threads = config["threads"] shell: """ @@ -38,12 +38,12 @@ rule circlator_localassembly: rule circlator_merge: input: - assembly = 'results/{sample}/assembly.fasta', - localassembly = "results/{sample}/circlator/03.assemble/assembly.fasta" - output: "results/{sample}/circlator/04.merge.fasta" + assembly = config["outdir"] + "/{sample}/assembly.fasta", + localassembly = config["outdir"] + "/{sample}/circlator/03.assemble/assembly.fasta" + output: config["outdir"] + "/{sample}/circlator/04.merge.fasta" params: - threads = config["threads"], - output_pref = "results/{sample}/circlator/04.merge" + threads = config["threads_per_job"], + output_pref = config["outdir"] + "/{sample}/circlator/04.merge" shell: """ circlator merge {input.assembly} {input.localassembly} {params.output_pref} --threads {params.threads} @@ -51,10 +51,10 @@ rule circlator_merge: """ rule circlator_clean: - input: "results/{sample}/circlator/04.merge.fasta" - output: "results/{sample}/circlator/05.clean.fasta" + input: config["outdir"] + "/{sample}/circlator/04.merge.fasta" + output: config["outdir"] + "/{sample}/circlator/05.clean.fasta" params: - output_pref = "results/{sample}/circlator/05.clean" + output_pref = config["outdir"] + "/{sample}/circlator/05.clean" shell: """ circlator clean {input} {params.output_pref} @@ -62,10 +62,10 @@ rule circlator_clean: """ rule circlator_fixstart: - input: "results/{sample}/circlator/05.clean.fasta" - output: "results/{sample}/circlator/06.fixstart.fasta" + input: config["outdir"] + "/{sample}/circlator/05.clean.fasta" + output: config["outdir"] + "/{sample}/circlator/06.fixstart.fasta" params: - output_pref = "results/{sample}/circlator/06.fixstart" + output_pref = config["outdir"] + "/{sample}/circlator/06.fixstart" shell: """ circlator fixstart {input} {params.output_pref} @@ -74,12 +74,12 @@ rule circlator_fixstart: rule rename: input: - assembly = "results/{sample}/circlator/06.fixstart.fasta" + assembly = config["outdir"] + "/{sample}/circlator/06.fixstart.fasta" params: prefix = "{sample}", keep_intermediate = config["keep_intermediate"] - output: "results/{sample}/assembly.circularized.renamed.fasta" + output: config["outdir"] + "/{sample}/assembly.circularized.renamed.fasta" run: diff --git a/assembly/workflow/rules/cleanup.smk b/assembly/workflow/rules/cleanup.smk deleted file mode 100644 index d0d3b2a..0000000 --- a/assembly/workflow/rules/cleanup.smk +++ /dev/null @@ -1,7 +0,0 @@ - # Remove circlator and pilon intermediate results, which take up a lot of space - if [ {params.keep_intermediate} == "No" ]; then - - rm results/{params.prefix}/noduplicates.fastq.gz - rm -r results/{params.prefix}/pilon - rm -r results/{params.prefix}/circlator - fi \ No newline at end of file diff --git a/assembly/workflow/rules/common.smk b/assembly/workflow/rules/common.smk old mode 100644 new mode 100755 diff --git a/assembly/workflow/rules/mapreads.smk b/assembly/workflow/rules/mapreads.smk old mode 100644 new mode 100755 diff --git a/assembly/workflow/rules/readQC.smk b/assembly/workflow/rules/readQC.smk old mode 100644 new mode 100755 index 2c5b50b..da08f9a --- a/assembly/workflow/rules/readQC.smk +++ b/assembly/workflow/rules/readQC.smk @@ -1,9 +1,9 @@ rule longQC: input: lambda wildcards: samples[wildcards.sample].longread_fastq - output: "results/{sample}/longqc/QC_vals_longQC_sampleqc.json" + output: config["outdir"] + "/{sample}/longqc/QC_vals_longQC_sampleqc.json" params: - threads = config["threads"], - outfolder = "results/{sample}/longqc/" + threads = config["threads_per_job"], + outfolder = config["outdir"] + "/{sample}/longqc/" shell: """ diff --git a/assembly/workflow/rules/summarize.smk b/assembly/workflow/rules/summarize.smk old mode 100644 new mode 100755 index 5a3cb7d..c64e64a --- a/assembly/workflow/rules/summarize.smk +++ b/assembly/workflow/rules/summarize.smk @@ -1,6 +1,7 @@ + rule read_summary: - input: expand("results/{sample}/longqc/QC_vals_longQC_sampleqc.json", sample = samples.keys()) - output: "results/read_summaries.tsv" + input: expand("{outdir}/{sample}/longqc/QC_vals_longQC_sampleqc.json", outdir = config["outdir"], sample = samples.keys()) + output: config["outdir"] + "/read_summaries.tsv" run: import json @@ -51,14 +52,16 @@ rule read_summary: rule assembly_summaries: - input: expand("results/{sample}/assembly_info.txt", sample = samples.keys()) - output: "results/assembly_summaries.tsv" + input: expand("{outdir}/{sample}/assembly_info.txt", outdir = config["outdir"], sample = samples.keys()) + output: + summaries = config["outdir"] + "/assembly_summaries.tsv", + single_contig_assemblies = config["outdir"] + "/single_contig_samples.txt" run: header = "" - outfile = open(output[0], "w") + outfile = open(output.summaries, "w") - outfile_single_contigs = open("results/single_contig_samples.txt", "w") + outfile_single_contigs = open(output.single_contig_assemblies, "w") for FILE in input: diff --git a/fig.png b/fig.png deleted file mode 100644 index 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mode 100644 new mode 100755 diff --git a/variantcalling/workflow/Snakefile b/variantcalling/workflow/Snakefile old mode 100644 new mode 100755 diff --git a/variantcalling/workflow/scripts/combine_assemblies.py b/variantcalling/workflow/scripts/combine_assemblies.py old mode 100644 new mode 100755 -- GitLab