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Generate reports per run, per project and per lane #13

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May 30, 2024
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a31040e
Generate reports per lane, group and rundir
Aratz Mar 18, 2024
0da5870
Improve formatting
Aratz Mar 28, 2024
d233d8f
Improve output sorting
Aratz Mar 28, 2024
307e43c
Use `group` instead of `project`
Aratz Apr 8, 2024
627cf94
Fix output channel
Aratz Apr 8, 2024
c9ba028
Fix linting
Aratz Apr 8, 2024
2cfc91d
Give credits back to NGI
Aratz Apr 8, 2024
fbfb02d
Fix file names
Aratz Apr 9, 2024
8a19929
Set up tests
Aratz May 3, 2024
eba628e
point test conf upstream
kedhammar May 3, 2024
23f69d1
project -> group
kedhammar May 3, 2024
1ebf3f1
project -> group
kedhammar May 3, 2024
b0bf471
make lane non-compulsory
kedhammar May 3, 2024
9e0eca3
remove unused file
kedhammar May 3, 2024
98e60bf
revamp nf-test, run once for each sequencing platform
kedhammar May 3, 2024
d95c660
Update modules and subworkflows
Aratz May 6, 2024
e0527cc
Fix multiqc extra files
Aratz May 7, 2024
42159a1
Add test snapshots
Aratz May 14, 2024
ef61f9f
Remove unused module configuration
Aratz May 14, 2024
51b01e9
Update usage docs and restore example samplesheet
Aratz May 14, 2024
7c7f31f
Update output docs
Aratz May 14, 2024
1c4f6e0
Update changelog
Aratz May 14, 2024
211bfaf
Update samplesheet in readme file
Aratz May 14, 2024
acc82d0
Merge remote-tracking branch 'origin/dev' into multiqc_multireport
Aratz May 14, 2024
df4e9cb
Run prettier
Aratz May 14, 2024
2303db9
Use testdata base path param in tests/MiSeq.main.nf.test
kedhammar May 16, 2024
1ea8ac0
Use testdata base path param in tests/NovaSeq6000.main.nf.test
kedhammar May 16, 2024
048765f
Use testdata base path param in tests/PromethION.main.nf.test
kedhammar May 16, 2024
4329bb9
Make the tests work with 'pipelines_testdata_base_path' parameter.
MatthiasZepper May 17, 2024
3ff8503
update snapshots, add nf-test.log to gitignore
kedhammar May 17, 2024
8ac4d76
visualize example run dir corresponsing to samplesheet
kedhammar May 17, 2024
84c9b3d
naming fixes
kedhammar May 20, 2024
aaf17b6
nf-core sync
kedhammar May 20, 2024
e6dfea9
nf-core fixes
kedhammar May 20, 2024
02affeb
Improve publishDir logic
Aratz May 30, 2024
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2 changes: 2 additions & 0 deletions .gitignore
Original file line number Diff line number Diff line change
Expand Up @@ -6,3 +6,5 @@ results/
testing/
testing*
*.pyc
.nf-test
.nf-test.log
2 changes: 2 additions & 0 deletions CHANGELOG.md
Original file line number Diff line number Diff line change
Expand Up @@ -9,6 +9,8 @@ Initial release of nf-core/seqinspector, created with the [nf-core](https://nf-c

### `Added`

- [#13](https://github.com/nf-core/seqinspector/pull/13) Generate reports per run, per project and per lane.

### `Fixed`

### `Dependencies`
Expand Down
15 changes: 4 additions & 11 deletions README.md
Original file line number Diff line number Diff line change
Expand Up @@ -39,26 +39,19 @@
> [!NOTE]
> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) with `-profile test` before running the workflow on actual data.

<!-- TODO nf-core: Describe the minimum required steps to execute the pipeline, e.g. how to prepare samplesheets.
Explain what rows and columns represent. For instance (please edit as appropriate):

First, prepare a samplesheet with your input data that looks as follows:

`samplesheet.csv`:

```csv
sample,fastq_1,fastq_2
CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
sample,lane,group,fastq_1,fastq_2,rundir
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what about an extra "individual" field for when you have multiple samples from the same patient (thinking cancer sample sarek style)?

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or is this what you mean by group?

CONTROL_REP1,1,GROUP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz,200624_A00834_0183_BHMTFYDRXX
```

Each row represents a fastq file (single-end) or a pair of fastq files (paired end).

-->

Now, you can run the pipeline using:

<!-- TODO nf-core: update the following command to include all required parameters for a minimal example -->

```bash
nextflow run nf-core/seqinspector \
-profile <docker/singularity/.../institute> \
Expand All @@ -80,11 +73,11 @@ For more details about the output files and reports, please refer to the

## Credits

nf-core/seqinspector was originally written by Adrien Coulier.
nf-core/seqinspector was originally written by the Swedish [@NationalGenomicsInfrastructure](https://github.com/NationalGenomicsInfrastructure/).

We thank the following people for their extensive assistance in the development of this pipeline:

<!-- TODO nf-core: If applicable, make list of people who have also contributed -->
- [@mahesh-panchal](https://github.com/mahesh-panchal)

## Contributions and Support

Expand Down
2 changes: 1 addition & 1 deletion assets/samplesheet.csv
Original file line number Diff line number Diff line change
@@ -1,3 +1,3 @@
sample,lane,project,fastq_1,fastq_2,rundir
sample,lane,group,fastq_1,fastq_2,rundir
SAMPLE_PAIRED_END,1,P001,/path/to/fastq/files/AEG588A1_S1_L002_R1_001.fastq.gz,/path/to/fastq/files/AEG588A1_S1_L002_R2_001.fastq.gz,/path/to/rundir
SAMPLE_SINGLE_END,2,P002,/path/to/fastq/files/AEG588A4_S4_L003_R1_001.fastq.gz,,/path/to/rundir
8 changes: 4 additions & 4 deletions assets/schema_input.json
Original file line number Diff line number Diff line change
Expand Up @@ -19,11 +19,11 @@
"errorMessage": "Lane ID must be a number",
"meta": ["lane"]
},
"project": {
"group": {
"type": "string",
"pattern": "^\\S+$",
"errorMessage": "Project ID cannot contain spaces",
"meta": ["project"]
"errorMessage": "Group ID cannot contain spaces",
"meta": ["group"]
},
"fastq_1": {
"type": "string",
Expand All @@ -47,7 +47,7 @@
"meta": ["rundir"]
}
},
"required": ["sample", "lane", "fastq_1"],
"required": ["sample", "fastq_1"],
"dependentRequired": {
"fastq_2": ["fastq_1"]
}
Expand Down
76 changes: 75 additions & 1 deletion conf/modules.config
Original file line number Diff line number Diff line change
Expand Up @@ -22,7 +22,7 @@ process {
ext.args = '--quiet'
}

withName: 'MULTIQC' {
withName: 'MULTIQC_GLOBAL' {
ext.args = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' }
publishDir = [
path: { "${params.outdir}/multiqc" },
Expand All @@ -31,4 +31,78 @@ process {
]
}

withName: 'MULTIQC_PER_LANE' {
ext.args = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' }
publishDir = [
path: { "${params.outdir}/multiqc/lanes" },
mode: params.publish_dir_mode,
saveAs: {
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filename ->
switch (filename) {
case 'versions.yml':
null
break
case ~/\[LANE:\d+\]_multiqc_(report\.html|plots|data)/:
def lane = (filename =~ /\[LANE:(\d+)\]_multiqc_(report\.html|plots|data)/)[0][1]
def new_filename = filename.replaceFirst(
"(?<prefix>.*)\\[LANE:${lane}\\]_(?<suffix>multiqc_(report\\.html|plots|data).*)",
'${prefix}${suffix}')
"L${lane}/${new_filename}"
break
default:
filename
}
}
]
}

withName: 'MULTIQC_PER_GROUP' {
ext.args = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' }
publishDir = [
path: { "${params.outdir}/multiqc/groups" },
mode: params.publish_dir_mode,
saveAs: {
filename ->
switch (filename) {
case 'versions.yml':
null
break
case ~/\[GROUP:.+\]_multiqc_(report\.html|plots|data)/:
def group = (filename =~ /\[GROUP:(.+)\]_multiqc_(report\.html|plots|data)/)[0][1]
def new_filename = filename.replaceFirst(
"(?<prefix>.*)\\[GROUP:${group}\\]_(?<suffix>multiqc_(report\\.html|plots|data).*)",
'${prefix}${suffix}')
"${group}/${new_filename}"
break
default:
filename
}
}
]
}

withName: 'MULTIQC_PER_RUNDIR' {
ext.args = { params.multiqc_title ? "--title \"$params.multiqc_title\"" : '' }
publishDir = [
path: { "${params.outdir}/multiqc/rundirss" },
mode: params.publish_dir_mode,
saveAs: {
filename ->
switch (filename) {
case 'versions.yml':
null
break
case ~/\[RUNDIR:.+\]_multiqc_(report\.html|plots|data)/:
def rundir = (filename =~ /\[RUNDIR:(.+)\]_multiqc_(report\.html|plots|data)/)[0][1]
def new_filename = filename.replaceFirst(
"(?<prefix>.*)\\[RUNDIR:${rundir}\\]_(?<suffix>multiqc_(report\\.html|plots|data).*)",
'${prefix}${suffix}')
"${rundir}/${new_filename}"
break
default:
filename
}
}
]
}
}
25 changes: 23 additions & 2 deletions docs/output.md
Original file line number Diff line number Diff line change
Expand Up @@ -6,8 +6,6 @@ This document describes the output produced by the pipeline. Most of the plots a

The directories listed below will be created in the results directory after the pipeline has finished. All paths are relative to the top-level results directory.

<!-- TODO nf-core: Write this documentation describing your workflow's output -->

## Pipeline overview

The pipeline is built using [Nextflow](https://www.nextflow.io/) and processes data using the following steps:
Expand Down Expand Up @@ -48,6 +46,29 @@ The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They m
- `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
- `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
- `multiqc_plots/`: directory containing static images from the report in various formats.
- `lanes/` [1]
- `L1/`
- `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
- `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
- `multiqc_plots/`: directory containing static images from the report in various formats.
- `L2/`
- ...
- `groups/` [1]
- `GROUPNAME1/`
- `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
- `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
- `multiqc_plots/`: directory containing static images from the report in various formats.
- `GROUPNAME2/`
- ...
- `rundir/` [1]
- `RUNDIR1/`
- `multiqc_report.html`: a standalone HTML file that can be viewed in your web browser.
- `multiqc_data/`: directory containing parsed statistics from the different tools used in the pipeline.
- `multiqc_plots/`: directory containing static images from the report in various formats.
- `RUNDIR2/`
- ...

[1] These files will only be generated if `lane`, `group` or `rundir` were specified for some samples.

</details>

Expand Down
44 changes: 21 additions & 23 deletions docs/usage.md
Original file line number Diff line number Diff line change
Expand Up @@ -10,47 +10,45 @@

## Samplesheet input

You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below.
You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location.

```bash
--input '[path to samplesheet file]'
```

### Multiple runs of the same sample
### Full samplesheet

The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. The pipeline will concatenate the raw reads before performing any downstream analysis. Below is an example for the same sample sequenced across 3 lanes:
The following simple run dir structure...

```csv title="samplesheet.csv"
sample,fastq_1,fastq_2
CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
CONTROL_REP1,AEG588A1_S1_L003_R1_001.fastq.gz,AEG588A1_S1_L003_R2_001.fastq.gz
CONTROL_REP1,AEG588A1_S1_L004_R1_001.fastq.gz,AEG588A1_S1_L004_R2_001.fastq.gz
```
run_dir
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Why did you replace the exemplary filenames with this synthetic example? I think this may lead to confusion, because it may prompt people to tediously rename their files prior to a run. We should make clear that seqinspector takes the relevant information from the columns in the sample sheet and not suggest that the file names matter.

Also, judgemental adjectives like "simple" (or "difficult" etc.) should ideally be avoided in a README.

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Since @mahesh-panchal requested we visualize the directory structure, I thought it would be easier to connect the dots to the example samplesheet if all the file names in the dir also contained the information shown in the samplesheet.

I don't necessarily get the impression we are suggesting the files need to follow a particular naming convention by showing an example that is as informative as possible, but I don't feel too strongly about it.

├── sample1_lane1_group1_r1.fq.gz
├── sample2_lane1_group1_r1.fq.gz
├── sample3_lane2_group2_r1.fq.gz
└── sample4_lane2_group3_r1.fq.gz
```

### Full samplesheet

The pipeline will auto-detect whether a sample is single- or paired-end using the information provided in the samplesheet. The samplesheet can have as many columns as you desire, however, there is a strict requirement for the first 3 columns to match those defined in the table below.

A final samplesheet file consisting of both single- and paired-end data may look something like the one below. This is for 6 samples, where `TREATMENT_REP3` has been sequenced twice.
...would be represented in the following samplesheet (shown as .tsv for readability)

```csv title="samplesheet.csv"
sample,fastq_1,fastq_2
CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
CONTROL_REP2,AEG588A2_S2_L002_R1_001.fastq.gz,AEG588A2_S2_L002_R2_001.fastq.gz
CONTROL_REP3,AEG588A3_S3_L002_R1_001.fastq.gz,AEG588A3_S3_L002_R2_001.fastq.gz
TREATMENT_REP1,AEG588A4_S4_L003_R1_001.fastq.gz,
TREATMENT_REP2,AEG588A5_S5_L003_R1_001.fastq.gz,
TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz,
TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz,
sample lane group fastq_1 fastq_2 rundir
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Do you think that the order of the columns is advisable like this? Intuitively, I would have put all categorical variables together at the end, so that additional columns can be added easily later, if required e.g. by other sequencing technologies.

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I don't have a strong opinion on this, and I feel this deserves to be discussed in a new issue/pr. This pr was not meant to change the input format. This commit to usage.md just fixes the documentation so that it's up to date with what the format actually is.

sample1 1 group1 path/to/run_dir/sample1_lane1_group1_r1.fq.gz path/to/run_dir
sample2 1 group1 path/to/run_dir/sample2_lane1_group1_r1.fq.gz path/to/run_dir
sample3 2 group2 path/to/run_dir/sample3_lane2_group2_r1.fq.gz path/to/run_dir
sample4 2 group3 path/to/run_dir/sample4_lane2_group3_r1.fq.gz path/to/run_dir

```

| Column | Description |
| --------- | -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- |
| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). |
| `lane` | Lane where the sample was processed on an Illumina instrument (optional). |
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| `group` | Group the sample belongs too, useful when several groups are pooled together (optional). |
| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". |
| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". |
| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz" (optional). |
| `rundir` | Path to the runfolder containing extra information about the sequencing run (optional) . |

An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline.
Another [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline.

## Running the pipeline

Expand Down
7 changes: 5 additions & 2 deletions main.nf
Original file line number Diff line number Diff line change
Expand Up @@ -58,7 +58,10 @@ workflow NFCORE_SEQINSPECTOR {
)

emit:
multiqc_report = SEQINSPECTOR.out.multiqc_report // channel: /path/to/multiqc_report.html
global_report = SEQINSPECTOR.out.global_report // channel: /path/to/multiqc_report.html
lane_reports = SEQINSPECTOR.out.lane_reports // channel: /path/to/multiqc_report.html
group_reports = SEQINSPECTOR.out.group_reports // channel: /path/to/multiqc_report.html
rundir_report = SEQINSPECTOR.out.rundir_reports // channel: /path/to/multiqc_report.html

}
/*
Expand Down Expand Up @@ -101,7 +104,7 @@ workflow {
params.outdir,
params.monochrome_logs,
params.hook_url,
NFCORE_SEQINSPECTOR.out.multiqc_report
NFCORE_SEQINSPECTOR.out.global_report,
)
}

Expand Down
8 changes: 8 additions & 0 deletions nf-test.config
Original file line number Diff line number Diff line change
@@ -0,0 +1,8 @@
config {

testsDir "tests"
workDir ".nf-test"
configFile "tests/nextflow.config"
profile "test,docker"

}
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Original file line number Diff line number Diff line change
Expand Up @@ -84,7 +84,7 @@ workflow PIPELINE_INITIALISATION {
.fromSamplesheet("input") // Validates samplesheet against $projectDir/assets/schema_input.json. Path to validation schema is defined by $projectDir/nextflow_schema.json
.map {
meta, fastq_1, fastq_2 ->
def id_string = "${meta.sample}_${meta.project ?: "ungrouped"}_${meta.lane}"
def id_string = "${meta.sample}_${meta.group ?: "ungrouped"}_${meta.lane}"
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Doesn't lane need a default value too if it's not required?

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It's been removed from required.

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😳 Have I then been reviewing the wrong/outdated version of this PR all the time? Because I ran gh pr 13 checkout and it is still in there for me locally ?!?

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I removed it so we would be able to run on sequencing platforms without lanes, e.g. ONT.

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Sure, but what about the other paths, e.g. channel where meta.group has a setting, but meta.lane has nothing (and the filter is on meta.group)?
The name will include null in it.

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Could you point out where this would be an issue?

I'll note that I don't mind re-working this code into something more explicit, I simply lack the know-how as of now 😆

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I guess the question is, is id used for anything important: For example with the Promethion test, the csv looks like:

sample,lane,group,fastq_1,fastq_2,rundir
hg001,,r10p41_e8p2_human_runs_jkw,https://github.com/nf-core/test-datasets/raw/seqinspector/testdata/PromethION/20230505_1857_1B_PAO99309_94e07fab/fastq_pass/PAO99309_pass__94e07fab_c3641428_1.fastq.gz,,

and then the id string should be: hg001_r10p41_e8p2_human_runs_jkw_null. Does it matter that this is the case?
When you're grouping the files by group r10p41_e8p2_human_runs_jkw I guess lane information is not needed at all downstream of this?

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Thanks for clarifying! From what I remember of the initial meeting, the id was intended as simply a concatenation of fields to ensure uniqueness within the pipeline run. This is still ensured even if some of the fields are null, right?

Intuitively I think having a consistent way to generate the id that sometimes contains null is preferable to setting up different conventions for generating it across different sequencing platforms.

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Wouldn't it be easier to use the user-provided sample column? Could be potentially combined with a short uuid or hash to be unique in case we have samples that extend over multiple input files?

def updated_meta = meta + [ id: id_string ]
if (!fastq_2) {
return [ updated_meta.id, updated_meta + [ single_end:true ], [ fastq_1 ] ]
Expand All @@ -101,7 +101,6 @@ workflow PIPELINE_INITIALISATION {
// meta, fastqs ->
// return [ meta, fastqs.flatten() ]
// }
.view()
.set { ch_samplesheet }

emit:
Expand Down
40 changes: 40 additions & 0 deletions tests/MiSeq.main.nf.test
Original file line number Diff line number Diff line change
@@ -0,0 +1,40 @@
nextflow_pipeline {

name "Test Workflow main.nf on MiSeq data"
script "../main.nf"
tag "seqinspector"
tag "PIPELINE"

test("MiSeq data test") {

when {
config "./MiSeq.main.nf.test.config"
params {
outdir = "$outputDir"
}
}

then {
assertAll(
{ assert workflow.success },
{ assert snapshot(
path("$outputDir/multiqc/lanes/L1/multiqc_data/multiqc_citations.txt"),
path("$outputDir/multiqc/lanes/L1/multiqc_data/multiqc_fastqc.txt"),
path("$outputDir/multiqc/lanes/L1/multiqc_data/multiqc_general_stats.txt"),
path("$outputDir/multiqc/lanes/L1/multiqc_data/multiqc_software_versions.txt"),

path("$outputDir/multiqc/groups/P001/multiqc_data/multiqc_citations.txt"),
path("$outputDir/multiqc/groups/P001/multiqc_data/multiqc_fastqc.txt"),
path("$outputDir/multiqc/groups/P001/multiqc_data/multiqc_general_stats.txt"),
path("$outputDir/multiqc/groups/P001/multiqc_data/multiqc_software_versions.txt"),

path("$outputDir/multiqc/multiqc_data/multiqc_citations.txt"),
path("$outputDir/multiqc/multiqc_data/multiqc_fastqc.txt"),
path("$outputDir/multiqc/multiqc_data/multiqc_general_stats.txt"),
path("$outputDir/multiqc/multiqc_data/multiqc_software_versions.txt"),
).match()
}
)
}
}
}
7 changes: 7 additions & 0 deletions tests/MiSeq.main.nf.test.config
Original file line number Diff line number Diff line change
@@ -0,0 +1,7 @@
// Load the basic test config
includeConfig 'nextflow.config'

// Load the correct samplesheet for that test
params {
input = params.pipelines_testdata_base_path + 'seqinspector/testdata/MiSeq/samplesheet.csv'
}
19 changes: 19 additions & 0 deletions tests/MiSeq.main.nf.test.snap
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{
"MiSeq data test": {
"content": [
"multiqc_citations.txt:md5,4c806e63a283ec1b7e78cdae3a923d4f",
"multiqc_fastqc.txt:md5,692b8aed0614ed1655f2c1cbea1ba312",
"multiqc_general_stats.txt:md5,630167d67d3f92408cd1a04422c7196f",
"multiqc_software_versions.txt:md5,7452f1f7aae2a8a4066c2ef6cd5ceb95",
"multiqc_citations.txt:md5,4c806e63a283ec1b7e78cdae3a923d4f",
"multiqc_fastqc.txt:md5,692b8aed0614ed1655f2c1cbea1ba312",
"multiqc_general_stats.txt:md5,630167d67d3f92408cd1a04422c7196f",
"multiqc_software_versions.txt:md5,7452f1f7aae2a8a4066c2ef6cd5ceb95",
"multiqc_citations.txt:md5,4c806e63a283ec1b7e78cdae3a923d4f",
"multiqc_fastqc.txt:md5,692b8aed0614ed1655f2c1cbea1ba312",
"multiqc_general_stats.txt:md5,630167d67d3f92408cd1a04422c7196f",
"multiqc_software_versions.txt:md5,7452f1f7aae2a8a4066c2ef6cd5ceb95"
],
"timestamp": "2024-05-30T13:14:20.263485"
}
}
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