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1 change: 0 additions & 1 deletion .editorconfig
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[*.{md,yml,yaml,html,css,scss,js,r,rmd,R,Rmd}]
indent_size = 2
trim_trailing_whitespace = false

# These files are edited and tested upstream in nf-core/modules
[/modules/nf-core/**]
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12 changes: 7 additions & 5 deletions .nf-core.yml
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lint:
nextflow_config:
- manifest.homePage
files_exist:
- conf/igenomes.config
- conf/igenomes_ignored.config
- assets/multiqc_config.yml
- conf/igenomes.config
- conf/igenomes_ignored.config
- assets/multiqc_config.yml
files_unchanged:
- .github/CONTRIBUTING.md
- assets/email_template.html
- assets/sendmail_template.txt
- .github/CONTRIBUTING.md
- assets/sendmail_template.txt
template_strings:
- assets/assesspool_report.Rmd
multiqc_config: false
nf_core_version: 3.3.1
repository_type: pipeline
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4 changes: 2 additions & 2 deletions CHANGELOG.md
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# assessPool: Changelog
# nf-core/assesspool: Changelog

The format is based on [Keep a Changelog](https://keepachangelog.com/en/1.0.0/)
and this project adheres to [Semantic Versioning](https://semver.org/spec/v2.0.0.html).

## v1.0.0dev - [date]

Initial release of assessPool, created with the [nf-core](https://nf-co.re/) template.
Initial release of nf-core/assesspool, created with the [nf-core](https://nf-co.re/) template.

### `Added`

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32 changes: 0 additions & 32 deletions CITATIONS.md
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## Pipeline tools

- [PoPoolation2](https://sourceforge.net/p/popoolation2/wiki/Tutorial/)

> Kofler, R., Pandey, R. V., & Schlötterer, C. (2011). PoPoolation2: identifying differentiation between populations using sequencing of pooled DNA samples (Pool-Seq). Bioinformatics, 27(24), 3435-3436.

- [poolfstat](https://doi.org/10.1111/1755-0998.13557)

> Gautier, M., Vitalis, R., Flori, L., & Estoup, A. (2022). f‐Statistics estimation and admixture graph construction with Pool‐Seq or allele count data using the R package poolfstat. Molecular Ecology Resources, 22(4), 1394-1416.

- [grenedalf](https://github.com/lczech/grenedalf)

> Czech, L., Spence, J. P., & Expósito-Alonso, M. (2024). grenedalf: population genetic statistics for the next generation of pool sequencing. Bioinformatics, 40(8), btae508.

- [bcftools](https://samtools.github.io/bcftools/bcftools.html)

> Danecek, P., Bonfield, J. K., Liddle, J., Marshall, J., Ohan, V., Pollard, M. O., ... & Li, H. (2021). Twelve years of SAMtools and BCFtools. Gigascience, 10(2), giab008.

- [vcftools](https://vcftools.github.io/)

> Danecek, P., Auton, A., Abecasis, G., Albers, C. A., Banks, E., DePristo, M. A., ... & 1000 Genomes Project Analysis Group. (2011). The variant call format and VCFtools. Bioinformatics, 27(15), 2156-2158.

- [samtools](https://samtools.github.io/)

> Danecek, P., Bonfield, J. K., Liddle, J., Marshall, J., Ohan, V., Pollard, M. O., ... & Li, H. (2021). Twelve years of SAMtools and BCFtools. Gigascience, 10(2), giab008.

- [Rsamtools](https://www.bioconductor.org/packages/release/bioc/html/Rsamtools.html)

> Morgan M, Pagès H, Obenchain V, Hayden N (2024). _Rsamtools: Binary alignment (BAM), FASTA, variant call (BCF), and tabix
file import_. doi:10.18129/B9.bioc.Rsamtools <https://doi.org/10.18129/B9.bioc.Rsamtools>, R package version 2.22.0,
<https://bioconductor.org/packages/Rsamtools>.



## Software packaging/containerisation tools

- [Anaconda](https://anaconda.com)
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2 changes: 1 addition & 1 deletion LICENSE
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MIT License

Copyright (c) The assessPool team
Copyright (c) The nf-core/assesspool team

Permission is hereby granted, free of charge, to any person obtaining a copy
of this software and associated documentation files (the "Software"), to deal
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133 changes: 49 additions & 84 deletions README.md
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<h1>
<picture>
<source media="(prefers-color-scheme: dark)" srcset="docs/images/logo.png" >
<img height="150px" alt="assessPool" src="docs/images/logo.png" >
<source media="(prefers-color-scheme: dark)" srcset="docs/images/nf-core-assesspool_logo_dark.png">
<img alt="nf-core/assesspool" src="docs/images/nf-core-assesspool_logo_light.png">
</picture>
</h1>

[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A50.24.04.2-brightgreen.svg)](https://www.nextflow.io/)
[![GitHub Actions CI Status](https://github.com/nf-core/assesspool/actions/workflows/ci.yml/badge.svg)](https://github.com/nf-core/assesspool/actions/workflows/ci.yml)
[![GitHub Actions Linting Status](https://github.com/nf-core/assesspool/actions/workflows/linting.yml/badge.svg)](https://github.com/nf-core/assesspool/actions/workflows/linting.yml)[![AWS CI](https://img.shields.io/badge/CI%20tests-full%20size-FF9900?labelColor=000000&logo=Amazon%20AWS)](https://nf-co.re/assesspool/results)[![Cite with Zenodo](http://img.shields.io/badge/DOI-10.5281/zenodo.XXXXXXX-1073c8?labelColor=000000)](https://doi.org/10.5281/zenodo.XXXXXXX)
[![nf-test](https://img.shields.io/badge/unit_tests-nf--test-337ab7.svg)](https://www.nf-test.com)

[![Nextflow](https://img.shields.io/badge/version-%E2%89%A524.04.2-green?style=flat&logo=nextflow&logoColor=white&color=%230DC09D&link=https%3A%2F%2Fnextflow.io)](https://www.nextflow.io/)
[![nf-core template version](https://img.shields.io/badge/nf--core_template-3.3.1-green?style=flat&logo=nfcore&logoColor=white&color=%2324B064&link=https%3A%2F%2Fnf-co.re)](https://github.com/nf-core/tools/releases/tag/3.3.1)
[![run with conda](http://img.shields.io/badge/run%20with-conda-3EB049?labelColor=000000&logo=anaconda)](https://docs.conda.io/en/latest/)
[![run with docker](https://img.shields.io/badge/run%20with-docker-0db7ed?labelColor=000000&logo=docker)](https://www.docker.com/)
[![run with singularity](https://img.shields.io/badge/run%20with-singularity-1d355c.svg?labelColor=000000)](https://sylabs.io/docs/)
[![Launch on Seqera Platform](https://img.shields.io/badge/Launch%20%F0%9F%9A%80-Seqera%20Platform-%234256e7)](https://cloud.seqera.io/launch?pipeline=https://github.com/nf-core/assesspool)

[![Get help on Slack](http://img.shields.io/badge/slack-nf--core%20%23assesspool-4A154B?labelColor=000000&logo=slack)](https://nfcore.slack.com/channels/assesspool)[![Follow on Bluesky](https://img.shields.io/badge/bluesky-%40nf__core-1185fe?labelColor=000000&logo=bluesky)](https://bsky.app/profile/nf-co.re)[![Follow on Mastodon](https://img.shields.io/badge/mastodon-nf__core-6364ff?labelColor=FFFFFF&logo=mastodon)](https://mstdn.science/@nf_core)[![Watch on YouTube](http://img.shields.io/badge/youtube-nf--core-FF0000?labelColor=000000&logo=youtube)](https://www.youtube.com/c/nf-core)

## Introduction

**assessPool** is a population genetics analysis pipeline designed for pooled sequencing runs (pool-seq). Starting from raw genomic variants (VCF or sync format), assessPool performs the following operations:

* Filters SNPs based on adjustable criteria with suggestions for pooled data
* Calculates population genetic statistics using [PoPoolation2](https://sourceforge.net/p/popoolation2/wiki/Main/), [poolfstat](https://doi.org/10.1111/1755-0998.13557), and/or [grenedalf](https://github.com/lczech/grenedalf).
* Generates an HTML report including visualizations of population genetic statistics
* Outputs results in tabular format for downstream analyses

Required inputs are a variant description file (sync or VCF) and a reference assembly (FASTA). These can be output from any number of reduced representation data processing pipelines (e.g., [grenepipe](https://github.com/moiexpositoalonsolab/grenepipe), [dDocent](https://ddocent.com/), etc.).

Major pipeline operations:

1. Import, index, and/or compress variant description and reference
1. Perform stepwise filtering to determine effects of individual filter options (count lost loci):
1. Min/max read depth
1. Minor allele count
1. Hardy-Weinberg equilibrium cutoff
1. Missing data
1. Allele length
1. Quality:depth ratio
1. Minimum read quality
1. Variant type
1. Mispaired read likelihood
1. Alternate observations
1. Mapping quality
1. Mapping ratio
1. Overall depth
1. Number of pools with data
1. Read balance
1. Variant thinning
1. Perform cumulative filtering (for VCF input)
1. Generate sync files
1. Unified (all pools)
1. Split pairwise
1. Generate allele frequency table
1. Calculate F<sub>st</sub>
1. PoPoolation2
1. &#123;poolfstat&#125;
1. grenedalf
1. Calculate Fisher's exact test for individual SNPs
1. PoPoolation2
1. assessPool native
1. Join frequency data to F<sub>st</sub> results
1. Extract contigs containing (user-configurable) strongly-differentiated loci
1. Create HTML report
1. Save all output data in tabular format for downstream analysis
**nf-core/assesspool** is a bioinformatics pipeline that ...

<!-- TODO nf-core:
Complete this sentence with a 2-3 sentence summary of what types of data the pipeline ingests, a brief overview of the
major pipeline sections and the types of output it produces. You're giving an overview to someone new
to nf-core here, in 15-20 seconds. For an example, see https://github.com/nf-core/rnaseq/blob/master/README.md#introduction
-->

<!-- TODO nf-core: Include a figure that guides the user through the major workflow steps. Many nf-core
workflows use the "tube map" design for that. See https://nf-co.re/docs/guidelines/graphic_design/workflow_diagrams#examples for examples. -->
<!-- TODO nf-core: Fill in short bullet-pointed list of the default steps in the pipeline -->

## Usage

> [!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.

First, prepare a sample sheet with your input data that looks as follows:
<!-- 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
project,input,vcf_index,reference,pools,pool_sizes
poolseq_test,data/pools.vcf.gz,data/pools.vcf.gz.tbi,data/ref.fasta,,"35,38,22,52,17,19"
sample,fastq_1,fastq_2
CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
```

Each row represents a complete pool-seq project or experiment. Column descriptions:
Each row represents a fastq file (single-end) or a pair of fastq files (paired end).

`project` (required): A brief unique identifier for this pool-seq project
`input` (required): Variant description file in sync or VCF format (optionally compressed using `bgzip`)
`vcf_index` (optional): TABIX-format index of the input VCF file (generated if not supplied)
`reference` (required): Reference assembly (FASTA)
`pools` (optional): Comma-separated list of pool names (will replace any names in the input file)
`pool_sizes` (required): Number of individuals in each pool. Either a single number for uniform pool sizes or a comma-separated list of sizes for each pool.


Required columns are `project`, `input`, `reference`, and `pool_sizes`.
-->

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 tobodev/assesspool \
nextflow run nf-core/assesspool \
-profile <docker/singularity/.../institute> \
--input samplesheet.csv \
--outdir <OUTDIR>
Expand All @@ -96,47 +67,41 @@ nextflow run tobodev/assesspool \
> [!WARNING]
> Please provide pipeline parameters via the CLI or Nextflow `-params-file` option. Custom config files including those provided by the `-c` Nextflow option can be used to provide any configuration _**except for parameters**_; see [docs](https://nf-co.re/docs/usage/getting_started/configuration#custom-configuration-files).

For more details and further functionality, please refer to the [usage documentation](docs/usage.md) and the [parameter documentation](parameters.md).

## Testing the pipeline
assessPool comes with two built-in profiles that allow the user to test the pipeline with a fully-functional input dataset. These profiles (whose descriptions can be found in [conf/test.confg](conf/test.config) and [conf/test_full.config](conf/test_full.config)) will run assessPool with either a full or reduced dataset of SNPs sequenced from wild populations of the coral *Montipora capitata*. Pipeline tests can be run by passing either `test` or `test_full` to the `-profile` option, along with a software/container management subsystem. For example, using singularity:

```
nextflow run tobodev/assesspool \
-profile test,singularity
```
or
```
nextflow run tobodev/assesspool \
-profile test_full,singularity
```
For more details and further functionality, please refer to the [usage documentation](https://nf-co.re/assesspool/usage) and the [parameter documentation](https://nf-co.re/assesspool/parameters).

## Pipeline output

Results of an example test run with a full size dataset can be found [here](https://tobodev.github.io/assesspool/).
To see the results of an example test run with a full size dataset refer to the [results](https://nf-co.re/assesspool/results) tab on the nf-core website pipeline page.
For more details about the output files and reports, please refer to the
[output documentation](docs/output.md).
[output documentation](https://nf-co.re/assesspool/output).

## Credits

assessPool was originally written by Evan B Freel, Emily E Conklin, Mykle L Hoban, Derek W Kraft, Jonathan L Whitney, Ingrid SS Knapp, Zac H Forsman, Robert J Toonen.
nf-core/assesspool was originally written by Evan B Freel, Emily E Conklin, Mykle L Hoban, Derek W Kraft, Jonathan L Whitney, Ingrid SS Knapp, Zac H Forsman, Robert J Toonen.

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

Richard Coleman, &#x02bb;Ale&#x02bb;alani Dudoit, and Cataixa López, who used assessPool during development and helped identify issues and suggest key feature improvements. We would also like to thank Iliana Baums, Tanya Beirne, Dave Carlon, Greg Conception, Matt Craig, Jeff Eble, Scott Godwin, Matt Iacchei, Frederique Kandel, Steve Karl, Jim Maragos, Bob Moffitt, Joe O'Malley, Lawrie Provost, Jennifer Salerno, Derek Skillings, Michael Stat, Ben Wainwright, and Kim Weersing, for their efforts in sample collection.
<!-- TODO nf-core: If applicable, make list of people who have also contributed -->

## Contributions and Support

If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md).

For further information or help, don't hesitate to get in touch on the [Slack `#assesspool` channel](https://nfcore.slack.com/channels/assesspool) (you can join with [this invite](https://nf-co.re/join/slack)).

## Citations

A list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.
<!-- TODO nf-core: Add citation for pipeline after first release. Uncomment lines below and update Zenodo doi and badge at the top of this file. -->
<!-- If you use nf-core/assesspool for your analysis, please cite it using the following doi: [10.5281/zenodo.XXXXXX](https://doi.org/10.5281/zenodo.XXXXXX) -->

<!-- TODO nf-core: Add bibliography of tools and data used in your pipeline -->

An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file.

This pipeline uses code and infrastructure developed and maintained by the [nf-core](https://nf-co.re) community, reused here under the [MIT license](https://github.com/nf-core/tools/blob/master/LICENSE).
You can cite the `nf-core` publication as follows:

> The nf-core framework for community-curated bioinformatics pipelines.
> **The nf-core framework for community-curated bioinformatics pipelines.**
>
> Philip Ewels, Alexander Peltzer, Sven Fillinger, Harshil Patel, Johannes Alneberg, Andreas Wilm, Maxime Ulysse Garcia, Paolo Di Tommaso & Sven Nahnsen.
>
> Nat Biotechnol. 2020 Feb 13. doi: 10.1038/s41587-020-0439-x.
> _Nat Biotechnol._ 2020 Feb 13. doi: [10.1038/s41587-020-0439-x](https://dx.doi.org/10.1038/s41587-020-0439-x).
2 changes: 1 addition & 1 deletion assets/adaptivecard.json
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"weight": "Bolder",
"color": "<% if (success) { %>Good<% } else { %>Attention<%} %>",
"text": "assessPool v${version} - ${runName}",
"text": "nf-core/assesspool v${version} - ${runName}",
"wrap": true
},
{
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