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8 | 8 | [](https://sylabs.io/docs/) |
9 | 9 | [](https://tower.nf/launch?pipeline=https://github.com/nf-core/rnaseq) |
10 | 10 |
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11 | | -[](https://nfcore.slack.com/channels/rnaseq)[](https://twitter.com/nf_core)[](https://www.youtube.com/c/nf-core) |
| 11 | +[](https://nfcore.slack.com/channels/rnaseq)[](https://twitter.com/nf_core)[](https://mstdn.science/@nf_core)[](https://www.youtube.com/c/nf-core) |
12 | 12 |
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13 | 13 | ## Introduction |
14 | 14 |
|
15 | | -<!-- TODO nf-core: Write a 1-2 sentence summary of what data the pipeline is for and what it does --> |
| 15 | +**nf-core/rnaseq** is a bioinformatics pipeline that ... |
16 | 16 |
|
17 | | -**nf-core/rnaseq** is a bioinformatics best-practice analysis pipeline for RNA sequencing analysis pipeline for gene/isoform quantification and extensive quality control.. |
18 | | - |
19 | | -The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It uses Docker/Singularity containers making installation trivial and results highly reproducible. The [Nextflow DSL2](https://www.nextflow.io/docs/latest/dsl2.html) implementation of this pipeline uses one container per process which makes it much easier to maintain and update software dependencies. Where possible, these processes have been submitted to and installed from [nf-core/modules](https://github.com/nf-core/modules) in order to make them available to all nf-core pipelines, and to everyone within the Nextflow community! |
20 | | - |
21 | | -<!-- TODO nf-core: Add full-sized test dataset and amend the paragraph below if applicable --> |
22 | | - |
23 | | -On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources.The results obtained from the full-sized test can be viewed on the [nf-core website](https://nf-co.re/rnaseq/results). |
24 | | - |
25 | | -## Pipeline summary |
| 17 | +<!-- TODO nf-core: |
| 18 | + Complete this sentence with a 2-3 sentence summary of what types of data the pipeline ingests, a brief overview of the |
| 19 | + major pipeline sections and the types of output it produces. You're giving an overview to someone new |
| 20 | + to nf-core here, in 15-20 seconds. For an example, see https://github.com/nf-core/rnaseq/blob/master/README.md#introduction |
| 21 | +--> |
26 | 22 |
|
| 23 | +<!-- TODO nf-core: Include a figure that guides the user through the major workflow steps. Many nf-core |
| 24 | + workflows use the "tube map" design for that. See https://nf-co.re/docs/contributing/design_guidelines#examples for examples. --> |
27 | 25 | <!-- TODO nf-core: Fill in short bullet-pointed list of the default steps in the pipeline --> |
28 | 26 |
|
29 | 27 | 1. Read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) |
30 | 28 | 2. Present QC for raw reads ([`MultiQC`](http://multiqc.info/)) |
31 | 29 |
|
32 | | -## Quick Start |
| 30 | +## Usage |
| 31 | + |
| 32 | +> **Note** |
| 33 | +> If you are new to Nextflow and nf-core, please refer to [this page](https://nf-co.re/docs/usage/installation) on how |
| 34 | +> to set-up Nextflow. Make sure to [test your setup](https://nf-co.re/docs/usage/introduction#how-to-run-a-pipeline) |
| 35 | +> with `-profile test` before running the workflow on actual data. |
| 36 | +
|
| 37 | +<!-- TODO nf-core: Describe the minimum required steps to execute the pipeline, e.g. how to prepare samplesheets. |
| 38 | + Explain what rows and columns represent. For instance (please edit as appropriate): |
| 39 | +
|
| 40 | +First, prepare a samplesheet with your input data that looks as follows: |
33 | 41 |
|
34 | | -1. Install [`Nextflow`](https://www.nextflow.io/docs/latest/getstarted.html#installation) (`>=22.10.1`) |
| 42 | +`samplesheet.csv`: |
35 | 43 |
|
36 | | -2. Install any of [`Docker`](https://docs.docker.com/engine/installation/), [`Singularity`](https://www.sylabs.io/guides/3.0/user-guide/) (you can follow [this tutorial](https://singularity-tutorial.github.io/01-installation/)), [`Podman`](https://podman.io/), [`Shifter`](https://nersc.gitlab.io/development/shifter/how-to-use/) or [`Charliecloud`](https://hpc.github.io/charliecloud/) for full pipeline reproducibility _(you can use [`Conda`](https://conda.io/miniconda.html) both to install Nextflow itself and also to manage software within pipelines. Please only use it within pipelines as a last resort; see [docs](https://nf-co.re/usage/configuration#basic-configuration-profiles))_. |
| 44 | +```csv |
| 45 | +sample,fastq_1,fastq_2 |
| 46 | +CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz |
| 47 | +``` |
37 | 48 |
|
38 | | -3. Download the pipeline and test it on a minimal dataset with a single command: |
| 49 | +Each row represents a fastq file (single-end) or a pair of fastq files (paired end). |
39 | 50 |
|
40 | | - ```bash |
41 | | - nextflow run nf-core/rnaseq -profile test,YOURPROFILE --outdir <OUTDIR> |
42 | | - ``` |
| 51 | +--> |
43 | 52 |
|
44 | | - Note that some form of configuration will be needed so that Nextflow knows how to fetch the required software. This is usually done in the form of a config profile (`YOURPROFILE` in the example command above). You can chain multiple config profiles in a comma-separated string. |
| 53 | +Now, you can run the pipeline using: |
45 | 54 |
|
46 | | - > - The pipeline comes with config profiles called `docker`, `singularity`, `podman`, `shifter`, `charliecloud` and `conda` which instruct the pipeline to use the named tool for software management. For example, `-profile test,docker`. |
47 | | - > - Please check [nf-core/configs](https://github.com/nf-core/configs#documentation) to see if a custom config file to run nf-core pipelines already exists for your Institute. If so, you can simply use `-profile <institute>` in your command. This will enable either `docker` or `singularity` and set the appropriate execution settings for your local compute environment. |
48 | | - > - If you are using `singularity`, please use the [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use) command to download images first, before running the pipeline. Setting the [`NXF_SINGULARITY_CACHEDIR` or `singularity.cacheDir`](https://www.nextflow.io/docs/latest/singularity.html?#singularity-docker-hub) Nextflow options enables you to store and re-use the images from a central location for future pipeline runs. |
49 | | - > - If you are using `conda`, it is highly recommended to use the [`NXF_CONDA_CACHEDIR` or `conda.cacheDir`](https://www.nextflow.io/docs/latest/conda.html) settings to store the environments in a central location for future pipeline runs. |
| 55 | +<!-- TODO nf-core: update the following command to include all required parameters for a minimal example --> |
50 | 56 |
|
51 | | -4. Start running your own analysis! |
| 57 | +```bash |
| 58 | +nextflow run nf-core/rnaseq \ |
| 59 | + -profile <docker/singularity/.../institute> \ |
| 60 | + --input samplesheet.csv \ |
| 61 | + --outdir <OUTDIR> |
| 62 | +``` |
52 | 63 |
|
53 | | - <!-- TODO nf-core: Update the example "typical command" below used to run the pipeline --> |
| 64 | +> **Warning:** |
| 65 | +> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those |
| 66 | +> provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; |
| 67 | +> see [docs](https://nf-co.re/usage/configuration#custom-configuration-files). |
54 | 68 |
|
55 | | - ```bash |
56 | | - nextflow run nf-core/rnaseq --input samplesheet.csv --outdir <OUTDIR> --genome GRCh37 -profile <docker/singularity/podman/shifter/charliecloud/conda/institute> |
57 | | - ``` |
| 69 | +For more details, please refer to the [usage documentation](https://nf-co.re/rnaseq/usage) and the [parameter documentation](https://nf-co.re/rnaseq/parameters). |
58 | 70 |
|
59 | | -## Documentation |
| 71 | +## Pipeline output |
60 | 72 |
|
61 | | -The nf-core/rnaseq pipeline comes with documentation about the pipeline [usage](https://nf-co.re/rnaseq/usage), [parameters](https://nf-co.re/rnaseq/parameters) and [output](https://nf-co.re/rnaseq/output). |
| 73 | +To see the the results of a test run with a full size dataset refer to the [results](https://nf-co.re/rnaseq/results) tab on the nf-core website pipeline page. |
| 74 | +For more details about the output files and reports, please refer to the |
| 75 | +[output documentation](https://nf-co.re/rnaseq/output). |
62 | 76 |
|
63 | 77 | ## Credits |
64 | 78 |
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