Metaphlan2 to use paired-end reads

tools/README.md

@@ -7,14 +7,14 @@ This directory contains all tool descriptions that can be imported and used with
 All tools are described in a `Snakefile` in a directory having its name.
 All Snakefiles try to respect some rules and best practices in their design:
 
-* Reference to options from a `config.yaml` file
+* Reference to options from a `config_example.yaml` file
 * Tool description
 
 ### Reference to options from `config.yaml` file
 
 First part correspond to all options that are set up from a `config.yaml` file. They all have the nomenclature `__TOOLNAME_variable`.
 
-Then in the YAML `config.yaml` file, you can set the variable as followed:
+Then in the YAML `config_example.yaml` file, you can set the variable as followed:
 
 ```yaml
 TOOLNAME:
@@ -31,17 +31,18 @@ config['TOOLNAME'].get('variable', 1)
 
 In order to ease the linking of tools in a workflow, every parts of the tool is described as followed:
 
-  * input with the nomenclature: `__TOOLNAME_input`
-  * output with the nomenclature: ` __TOOLNAME_output`
-  * params with different options that are described above
-  	* There is usually a `exec_command` to give the possibility to change the way the tool is called (locally installed, singularity ...)
-  	* There is usually a `options` to specify all other command line options. _You can still give a more detailed level of description for the options_
-  * the shell command
+* input with the nomenclature: `__TOOLNAME_input`
+* output with the nomenclature: ` __TOOLNAME_output`
+* params with different options that are described above
+  * There is usually a `exec_command` to give the possibility to change the way the tool is called (locally installed, singularity ...)
+  * There is usually a `options` to specify all other command line options. _You can still give a more detailed level of description for the options_
+* the shell command
 
 input and output are then set up in the workflow `Snakefile` that refer to the rule.
 Therefore the rules cannot be used directly.
 
+> **Info**: You have noticed the possibility to give `modules` this is dedicated to our HPC that have some tools accessible via `module`.
+
 ## Example
 
-The directory for metaphlan2 rules give some example on the way to call the rules as well as setting
-the parameters in the `config.yaml` file.
+To find out example, refer to the workflow based on this tool descriptions in the `/workflows/` section.

tools/metaphlan2/metaphlan2/paired/Snakefile

+__metaphlan2_exec_command = config.get('metaphlan2', {}).get('exec_command', 'metaphlan2.py')
+__metaphlan2_modules = config.get('metaphlan2', {}).get('modules')
+__metaphlan2_input_type = config['metaphlan2'].get('input_type', 'fastq')
+__metaphlan2_options = config.get('metaphlan2', {}).get('options', "")
+__metaphlan2_threads = config.get('metaphlan2', {}).get('threads', 1)
+
+
+rule metaphlan2_paired:
+    """
+    MetaPhlAn 2 can also natively handle paired-end metagenomes (but does not use the paired-end information),
+    and, more generally, metagenomes stored in multiple files (but you need to specify the --bowtie2out parameter):
+
+    $ metaphlan2.py metagenome_1.fastq,metagenome_2.fastq --bowtie2out metagenome.bowtie2.bz2 --nproc 5 
+                    --input_type fastq > profiled_metagenome.txt
+
+    """
+    input:
+        r1 = __metaphlan2_input_r1,
+        r2 = __metaphlan2_input_r2
+    output:
+        __metaphlan2_output
+    params:
+        exec_command = __metaphlan2_exec_command,
+        modules = __metaphlan2_modules,
+        input_type = __metaphlan2_input_type,
+        bowtie2out = "{output_dir}/{sample}.bowtie2.bz2".format(output_dir=__metaphlan2_output_dir, sample="{sample}"),
+        options = __metaphlan2_options
+    threads:
+        __metaphlan2_threads
+    run:
+        command = []
+        if params.modules:
+            command.append("module load {params.modules}")
+        command.append("{params.exec_command} --nproc {threads} --input_type {params.input_type} --bowtie2out {params.bowtie2out} {params.options} {input.r1},{input.r2} {output}")
+        shell(" && ".join(command))

tools/metaphlan2/metaphlan2/paired/config_example.yaml

+input_dir: data
+
+metaphlan2:
+  threads: 1
+  input_type: fastq
+  options: ""
+  pair_suffix: ""
+  exec_command: metaphlan2.py

tools/metaphlan2/metaphlan2/Snakefile → tools/metaphlan2/metaphlan2/single/Snakefile

 __metaphlan2_exec_command = config.get('metaphlan2', {}).get('exec_command', 'metaphlan2.py')
 __metaphlan2_modules = config.get('metaphlan2', {}).get('modules')
+__metaphlan2_input_type = config['metaphlan2'].get('input_type', 'fastq')
 __metaphlan2_options = config.get('metaphlan2', {}).get('options', "")
 __metaphlan2_threads = config.get('metaphlan2', {}).get('threads', 1)
 
+
 rule metaphlan2:
     input:
         __metaphlan2_input
@@ -12,6 +14,7 @@ rule metaphlan2:
         exec_command = __metaphlan2_exec_command,
         modules = __metaphlan2_modules,
         input_type = __metaphlan2_input_type,
+        bowtie2out = "{output_dir}/{sample}.bowtie2.bz2".format(output_dir=__metaphlan2_output_dir, sample="{sample}"),
         options = __metaphlan2_options
     threads:
         __metaphlan2_threads
@@ -19,5 +22,5 @@ rule metaphlan2:
         command = []
         if params.modules:
             command.append("module load {params.modules}")
-        command.append("{params.exec_command} --nproc {threads} --input_type {params.input_type} {params.options} {input} {output}")
+        command.append("{params.exec_command} --nproc {threads} --input_type {params.input_type} --bowtie2out {params.bowtie2out} {params.options} {input} {output}")
         shell(" && ".join(command))

tools/metaphlan2/metaphlan2/usage_example.rules

-"""
-This example would be used as followed:
-
-$ snakemake --snakefile metaphlan2.rules output/s01.txt 
-
-It requires the presence of data/s01.fastq.gz to work based on config file
-"""
-
-configfile: "config.yaml"
-
-__input_dir = config['input_dir']
-__main_output_dir = config.get('output_dir', 'output')
-
-# ---- Metaphlan2
-__metaphlan2_output_dir = __main_output_dir + "/metaphlan2"
-__metaphlan2_input_type = config['metaphlan2'].get('input_type', 'fastq')
-__metaphlan2_input = "{dir}/{sample}.{ext}".format(dir=__input_dir,
-                                                   sample="{sample}",
-                                                   ext=__metaphlan2_input_type + ".gz")
-__metaphlan2_output = "{dir}/{sample}.txt".format(dir=__metaphlan2_output_dir,
-                                                  sample="{sample}")
-include: "Snakefile"
-
-rule all:
-    input:
-        __metaphlan2_output

workflows/README.md

@@ -13,6 +13,8 @@ Therefore, workflows have the following parts:
 4. Set up of specific variables for the tools
 5. `rule all` to specify what is/are the specific file(s) expected from the workflow.
 
+> **Note**: These workflows are not made to be fully portable and reusable but at least give around 80% of the work done. You might need to do minor modifications. The idea being to have a copy of `Snakefile` workflow with its dedicated `config.yaml` file for each experiments. 
+
 ### Reference to options from `config.yaml` file
 
 First part correspond to all options that are set up from a `config.yaml` file. They all have the nomenclature `__TOOLNAME_variable`.

workflows/metaphlan2/README.md

+# Simple metaphlan2 workflows
+
+Workflows using metaphlan2 and simple visualization of the results.
+
+All examples presented were made for our TARS cluster system. This means you will be likely to find some
+absolute path into the `config.yaml` that you might not have access to.
+
+For every workflow, an example is provided and is based on the `config.yaml` file. Singularity images are necessary for these examples.

workflows/metaphlan2/paired_metaphlan2/Snakefile

+configfile: "config.yaml"
+
+# ==== Snakefile paths ==== 
+__metaphlan2_rules = config.get("snakefiles", {}).get("metaphlan2", "../../tools/metaphlan2/metaphlan2/Snakefile")
+__metaphlan2_merge_rules = config.get("snakefiles", {}).get("metaphlan2_merge", "../../tools/metaphlan2/metaphlan2_merge/Snakefile")
+__metaphlan2_heatmap_rules = config.get("snakefiles", {}).get("metaphlan2_heatmap", "../../tools/metaphlan2/metaphlan2_heatmap/Snakefile")
+__graphlan_from_metaphlan2_rules = config.get("snakefiles", {}).get("graphlan_from_metaphlan2", "../subworkflows/graphlan_from_metaphlan2/Snakefile")
+
+__input_dir = config['input_dir']
+__main_output_dir = config.get('output_dir', 'output')
+
+# ---- Metaphlan2
+__metaphlan2_suffix = config['metaphlan2'].get('pair_suffix', '')
+__metaphlan2_output_dir = __main_output_dir + "/metaphlan2"
+__metaphlan2_input_type = config['metaphlan2'].get('input_type', 'fastq')
+__metaphlan2_input_r1 = "{dir}/{sample}_R1{suffix}.{ext}".format(dir=__input_dir,
+                                                                 sample="{sample}",
+                                                                 suffix=__metaphlan2_suffix,
+                                                                 ext=__metaphlan2_input_type + ".gz")
+__metaphlan2_input_r2 = "{dir}/{sample}_R2{suffix}.{ext}".format(dir=__input_dir,
+                                                                 sample="{sample}",
+                                                                 suffix=__metaphlan2_suffix,
+                                                                 ext=__metaphlan2_input_type + ".gz")
+__metaphlan2_output = "{dir}/{sample}.txt".format(dir=__metaphlan2_output_dir,
+                                                  sample="{sample}")
+include: __metaphlan2_rules
+
+# ---- Metaphlan2 merge
+__metaphlan2_merge_output_dir = __main_output_dir + "/metaphlan2_merge"
+__metaphlan2_merge_output_file_name = config['metaphlan2_merge'].get('output_file_name',"merged_taxonomic_profiles.txt")
+__metaphlan2_merge_input = expand("{dir}/{sample}.txt".format(dir=__metaphlan2_output_dir,
+                                                              sample="{sample}"),
+                                  sample=config['samples'])
+__metaphlan2_merge_output = "{dir}/{file_name}".format(dir=__metaphlan2_merge_output_dir,
+                                                       file_name=__metaphlan2_merge_output_file_name) 
+include: __metaphlan2_merge_rules
+
+# ---- Metaphlan2 heatmap
+__metaphlan2_heatmap_output_dir = __main_output_dir + "/metaphlan2_heatmap"
+__metaphlan2_heatmap_output_file_name = config['metaphlan2_heatmap'].get('output_name',"heatmap.png")
+__metaphlan2_heatmap_input = __metaphlan2_merge_output
+__metaphlan2_heatmap_output = "{dir}/{file_name}".format(dir=__metaphlan2_heatmap_output_dir,
+                                                         file_name=__metaphlan2_heatmap_output_file_name) 
+include: __metaphlan2_heatmap_rules
+
+# ---- Graphlan Dendogram
+__graphlan_from_metaphlan2_output_dir = __main_output_dir + "/graphlan"
+__graphlan_from_metaphlan2_output_file_name =  config.get("graphlan_from_metaphlan2", {}).get('output_name',"dendogram.png")
+__graphlan_from_metaphlan2_input = __metaphlan2_merge_output
+__graphlan_from_metaphlan2_output = "{dir}/{file_name}".format(dir=__graphlan_from_metaphlan2_output_dir,
+                                                         file_name=__graphlan_from_metaphlan2_output_file_name)
+include: __graphlan_from_metaphlan2_rules
+
+rule all:
+    input:
+        heatmap = __metaphlan2_heatmap_output,
+        dendogram = __graphlan_from_metaphlan2_output
+

workflows/metaphlan2/paired_metaphlan2/config.yaml

+snakefiles:
+  metaphlan2: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2/paired/Snakefile
+  metaphlan2_merge: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2_merge/Snakefile
+  metaphlan2_heatmap: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2_heatmap/Snakefile
+  graphlan_from_metaphlan2: /pasteur/projets/policy01/Atm/snakemake/subworkflows/graphlan_from_metaphlan2/Snakefile
+
+samples:
+  - sample_1
+  - sample_2
+  - sample_2
+
+input_dir: /a/path/to/input/data
+output_dir: metaphlan2_output
+
+metaphlan2:
+  modules: singularity
+  threads: 4
+  input_type: fastq
+  pair_suffix: "_001"
+  options: --bowtie2db /pasteur/gaia/projets/p01/Atm/DBs/bowtie2/metaphlan2/
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.7.7_s3.2.1.simg metaphlan2.py
+
+metaphlan2_merge:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.7.7_s3.2.1.simg merge_metaphlan_tables.py
+
+metaphlan2_heatmap:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.6.0_s3.2.1.simg metaphlan_hclust_heatmap.py
+  output_name: snakemake_heatmap.png
+
+export2graphlan:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg export2graphlan.py
+  options: "--skip_rows 1,2 --most_abundant 100 --abundance_threshold 1 --least_biomarkers 10 --annotations 5,6 --external_annotations 7 --min_clade_size 1"
+
+graphlan_annotate:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg graphlan_annotate.py
+
+graphlan:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg graphlan.py
+  options: "--dpi 300 --external_legends"

workflows/simple_metaphlan2/Snakefile → workflows/metaphlan2/single_metaphlan2/Snakefile

 configfile: "config.yaml"
 
 # ==== Snakefile paths ==== 
-__metaphlan2_rules = config.get("snakefiles", {}).get("metaphlan2", "../../tools/metaphlan2/metaphlan2/Snakefile")
-__metaphlan2_merge_rules = config.get("snakefiles", {}).get("metaphlan2_merge", "../../tools/metaphlan2/metaphlan2_merge/Snakefile")
-__metaphlan2_heatmap_rules = config.get("snakefiles", {}).get("metaphlan2_heatmap", "../../tools/metaphlan2/metaphlan2_heatmap/Snakefile")
-__graphlan_from_metaphlan2_rules = config.get("snakefiles", {}).get("graphlan_from_metaphlan2", "../subworkflows/graphlan_from_metaphlan2/Snakefile")
+__metaphlan2_rules = config.get("snakefiles", {}).get("metaphlan2")
+__metaphlan2_merge_rules = config.get("snakefiles", {}).get("metaphlan2_merge")
+__metaphlan2_heatmap_rules = config.get("snakefiles", {}).get("metaphlan2_heatmap")
+__graphlan_from_metaphlan2_rules = config.get("snakefiles", {}).get("graphlan_from_metaphlan2")
 
 __input_dir = config['input_dir']
 __main_output_dir = config.get('output_dir', 'output')

workflows/metaphlan2/single_metaphlan2/config.yaml

+snakefiles:
+  metaphlan2: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2/single/Snakefile
+  metaphlan2_merge: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2_merge/Snakefile
+  metaphlan2_heatmap: /pasteur/projets/policy01/Atm/snakemake/tools/metaphlan2/metaphlan2_heatmap/Snakefile
+  graphlan_from_metaphlan2: /pasteur/projets/policy01/Atm/snakemake/subworkflows/graphlan_from_metaphlan2/Snakefile
+
+samples:
+  - sample_1
+  - sample_2
+  - sample_2
+
+input_dir: /a/path/to/input/data
+output_dir: metaphlan2_output
+
+metaphlan2:
+  modules: singularity
+  threads: 4
+  input_type: fastq
+  options: --bowtie2db /pasteur/gaia/projets/p01/Atm/DBs/bowtie2/metaphlan2/
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.7.7_s3.2.1.simg metaphlan2.py
+
+metaphlan2_merge:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.7.7_s3.2.1.simg merge_metaphlan_tables.py
+
+metaphlan2_heatmap:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/metaphlan2/from_docker/metaphlan2_2.6.0_s3.2.1.simg metaphlan_hclust_heatmap.py
+  output_name: snakemake_heatmap.png
+
+export2graphlan:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg export2graphlan.py
+  options: "--skip_rows 1,2 --most_abundant 100 --abundance_threshold 1 --least_biomarkers 10 --annotations 5,6 --external_annotations 7 --min_clade_size 1"
+
+graphlan_annotate:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg graphlan_annotate.py
+
+graphlan:
+  modules: singularity
+  exec_command: singularity exec --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/tools/graphlan/from_docker/graphlan_0.9.7_s3.2.1.simg graphlan.py
+  options: "--dpi 300 --external_legends"

workflows/simple_metaphlan2/README.md

-# Simple metaphlan2 workflow
-
-This is a simple example of a workflow using metaphlan2.
-
-## Example
-
-An example is provided and is based on the `config.yaml` file. Singularity images are necessary for this example as well as the samples in the appropriate folder.
\ No newline at end of file

workflows/simple_metaphlan2/config.yaml

-samples:
-  - SRS014459-Stool
-  - SRS014464-Anterior_nares
-  - SRS014470-Tongue_dorsum
-  - SRS014472-Buccal_mucosa
-  - SRS014476-Supragingival_plaque
-  - SRS014494-Posterior_fornix
-
-input_dir: /pasteur/homes/kehillio/Atm/kenzo/repo/workflow-benchmarking/data
-output_dir: 20190509_simplecase
-
-metaphlan2:
-  threads: 4
-  input_type: fasta
-  exec_command: singularity run --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/metaphlan2/metaphlan2.simg
-
-metaphlan2_merge:
-  exec_command: singularity run --app merge --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/metaphlan2/metaphlan2.simg 
-
-metaphlan2_heatmap:
-  exec_command: singularity run --app heatmap --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/metaphlan2/metaphlan2.simg
-  output_name: snakemake_heatmap.png
-
-export2graphlan:
-  exec_command: singularity run --app export2graphlan --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/graphlan/graphlan.simg
-  options: "--skip_rows 1,2 --most_abundant 100 --abundance_threshold 1 --least_biomarkers 10 --annotations 5,6 --external_annotations 7 --min_clade_size 1"
-
-graphlan_annotate:
-  exec_command: singularity run --app graphlan_annotate --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/graphlan/graphlan.simg
-
-graphlan:
-  exec_command: singularity run --bind /pasteur/ /pasteur/gaia/projets/p01/Atm/singularity/graphlan/graphlan.simg
-  options: "--dpi 300 --external_legends"
\ No newline at end of file