---
-title: " Analyzing multiple samples"
-teaching: 0
-exercises: 0
+title: "Analyzing Multiple Samples"
+teaching: 30
+exercises: 30
questions:
-- "Key question (FIXME)"
+- "How can you run the same workflow over multiple samples?"
objectives:
-- "First learning objective. (FIXME)"
+- "Modify the workflow to process multiple samples, then perform a joint analysis."
keypoints:
-- "First key point. Brief Answer to questions. (FIXME)"
+- "Separate the part of the workflow that you want to run multiple times into a subworkflow."
+- "Use a scatter step to run the subworkflow over a list of inputs."
+- "The result of a scatter is an array, which can be used in a combine step to get a single result."
---
-# Analyzing multiple samples
+In the previous lesson, we completed converting the function of the
+original source shell script into CWL. This lesson expands the scope
+by demonstrating what changes to make to the workflow to be able to
+analyze multiple samples in parallel.
-Analyzing a single sample is great, but in the real world you probably
-have a batch of samples that you need to analyze and then compare.
-
-### 1. Subworkflows
+# Subworkflows
In addition to running command line tools, a workflow step can also
execute another workflow.
-Let's copy "main.cwl" to "alignment.cwl".
+First, copy `main.cwl` to `alignment.cwl`.
+
+Next, open `main.cwl` for editing. We are going to replace the `steps` and `outputs` sections.
-Now, edit open "main.cwl" for editing. We are going to replace the `steps` and `outputs` sections.
+Remove all the steps and replace them with a single `alignment` step
+which invokes the `alignment.cwl` we just copied.
```
steps:
gtf: gtf
out: [qc_html, bam_sorted_indexed, featurecounts]
```
+{: .language-yaml }
-In the outputs section, all the output sources are from the alignment step:
+In the `outputs` section, all the output sources are from the alignment step:
```
outputs:
type: File
outputSource: alignment/featurecounts
```
+{: .language-yaml }
-We also need a little boilerplate to tell the workflow runner that we want to use subworkflows:
+We also need add "SubworkflowFeatureRequirement" to tell the workflow
+runner that we are using subworkflows:
```
requirements:
SubworkflowFeatureRequirement: {}
```
+{: .language-yaml }
+
+> ## Running the workflow
+>
+> Run this workflow. You should get exactly the same results as
+> before, as all we have done so far is to wrap the inner workflow with
+> an outer workflow.
+>
+{: .challenge }
-If you run this workflow, you will get exactly the same results as
-before, we've just wrapped the inner workflow with an outer workflow.
+> ## Part 1 solution
+> * <a href="../assets/answers/ep5/part1/main.cwl">main.cwl</a>
+> * <a href="../assets/answers/ep5/part1/alignment.cwl">alignment.cwl</a>
+> * <a href="../assets/answers/ep5/part1/featureCounts.cwl">featureCounts.cwl</a>
+{: .solution}
-### 2. Scattering
+# Scattering
-The wrapper lets us do something useful. We can modify the outer
-workflow to accept a list of files, and then invoke the inner workflow
-step for every one of those files. We will need to modify the
-`inputs`, `steps`, `outputs`, and `requirements` sections.
+The "wrapper" step lets us do something useful. We can modify the
+outer workflow to accept a list of files, and then invoke the inner
+workflow step for every one of those files. We will need to modify
+the `inputs`, `steps`, `outputs`, and `requirements` sections.
First we change the `fq` parameter to expect a list of files:
genome: Directory
gtf: File
```
+{: .language-yaml }
-Next, we add `scatter` to the alignment step. The means it will
-run `alignment.cwl` for each value in the list in the `fq` parameter.
+Next, we add `scatter` to the alignment step. The means we want to
+run run `alignment.cwl` for each value in the list in the `fq`
+parameter.
```
steps:
gtf: gtf
out: [qc_html, bam_sorted_indexed, featurecounts]
```
+{: .language-yaml }
Because the scatter produces multiple outputs, each output parameter
becomes a list as well:
type: File[]
outputSource: alignment/featurecounts
```
+{: .language-yaml }
-Finally, we need a little more boilerplate to tell the workflow runner
-that we want to use scatter:
+We also need add "ScatterFeatureRequirement" to tell the workflow
+runner that we are using scatter:
```
requirements:
SubworkflowFeatureRequirement: {}
ScatterFeatureRequirement: {}
```
+{: .language-yaml }
-### 3. Running with list inputs
+> ## Part 2 solution
+> * <a href="../assets/answers/ep5/part2/main.cwl">main.cwl</a>
+> * <a href="../assets/answers/ep5/part2/alignment.cwl">alignment.cwl</a>
+> * <a href="../assets/answers/ep5/part2/featureCounts.cwl">featureCounts.cwl</a>
+{: .solution}
+
+# Input parameter lists
The `fq` parameter needs to be a list. You write a list in yaml by
starting each list item with a dash. Example `main-input.yaml`
class: File
location: rnaseq/reference_data/chr1-hg19_genes.gtf
```
+{: .language-yaml }
-Now you can run the workflow the same way as in Lesson 2.
+> ## Running the workflow
+>
+> Run this workflow. You will now get results for each one of the
+> input fastq files.
+>
+{: .challenge }
-### 4. Combining results
+# Combining results
-Each instance of the alignment workflow produces its own featureCounts
-file. However, to be able to compare results easily, we need them a
-single file with all the results.
+Each instance of the alignment workflow produces its own
+`featurecounts.tsv` file. However, to be able to compare results
+easily, we would like single file with all the results.
-The easiest way to do this is to run `featureCounts` just once at the
-end of the workflow, with all the bam files listed on the command
-line.
+We can modify the workflow to run `featureCounts` once at the end of
+the workflow, taking all the bam files listed on the command line.
-We'll need to modify a few things.
+We will need to change a few things.
First, in `featureCounts.cwl` we need to modify it to accept either a
single bam file or list of bam files.
- File
- File[]
```
+{: .language-yaml }
Second, in `alignment.cwl` we need to remove the `featureCounts` step from alignment.cwl, as well as the `featurecounts` output parameter.
gtf: gtf
out: [featurecounts]
```
+{: .language-yaml }
Last, we modify the `featurecounts` output parameter. Instead of a
list of files produced by the `alignment` step, it is now a single
type: File
outputSource: featureCounts/featurecounts
```
-
-Run this workflow to get a single `featurecounts.tsv` file with a column for each bam file.
+{: .language-yaml }
+
+> ## Running the workflow
+>
+> Run this workflow. You will still have separate results from fastq
+> and and STAR, but now you will only have a single
+> `featurecounts.tsv` file with a column for each bam file.
+>
+{: .challenge }
+
+> ## Episode solution
+> * <a href="../assets/answers/ep5/part4/main.cwl">main.cwl</a>
+> * <a href="../assets/answers/ep5/part4/alignment.cwl">alignment.cwl</a>
+> * <a href="../assets/answers/ep5/part4/featureCounts.cwl">featureCounts.cwl</a>
+{: .solution}
projects / rnaseq-cwl-training.git / blobdiff