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--- | ||
title: genomic-medicine-sweden/nallo | ||
description: A bioinformatics analysis pipeline for long-reads from both PacBio and (targeted) ONT-data, focused on rare-disease. | ||
--- | ||
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## Introduction | ||
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**genomic-medicine-sweden/nallo** is a bioinformatics analysis pipeline for long-reads from both PacBio and (targeted) ONT-data, focused on rare-disease. Heavily influenced by best-practice pipelines such as [nf-core/sarek](https://nf-co.re/sarek), [nf-core/raredisease](https://nf-co.re/raredisease), [nf-core/nanoseq](https://github.com/nf-core/nanoseq), [PacBio Human WGS Workflow](https://github.com/PacificBiosciences/pb-human-wgs-workflow-snakemake), [epi2me-labs/wf-human-variation](https://github.com/epi2me-labs/wf-human-variation) and [brentp/rare-disease-wf](https://github.com/brentp/rare-disease-wf). | ||
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## Overview | ||
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<picture align="center"> | ||
<img alt="genomic-medicine-sweden/nallo workflow" src="docs/images/nallo_metromap.png"> | ||
</picture> | ||
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## Pipeline summary | ||
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##### QC | ||
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- Read QC with [FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/), [cramino](https://github.com/wdecoster/cramino) and [mosdepth](https://github.com/brentp/mosdepth) | ||
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##### Alignment & assembly | ||
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- Align reads to reference with [minimap2](https://github.com/lh3/minimap2) | ||
- Assemble (trio-binned) haploid genomes with [hifiasm](https://github.com/chhylp123/hifiasm) (HiFi only) | ||
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##### Variant calling | ||
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- Call SNVs & joint genotyping with [deepvariant](https://github.com/google/deepvariant) and [GLNexus](https://github.com/dnanexus-rnd/GLnexus) | ||
- Call SVs with [Severus](https://github.com/KolmogorovLab/Severus) or [Sniffles2](https://github.com/fritzsedlazeck/Sniffles) | ||
- Call CNVs with [HiFiCNV](https://github.com/PacificBiosciences/HiFiCNV) | ||
- Call tandem repeats with [TRGT](https://github.com/PacificBiosciences/trgt/tree/main) (HiFi only) | ||
- Call paralogous genes with [Paraphase](https://github.com/PacificBiosciences/paraphase) | ||
- Call variants from assembly with [dipcall](https://github.com/lh3/dipcall) (HiFi only) | ||
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##### Phasing and methylation | ||
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- Phase and haplotag reads with [LongPhase](https://github.com/twolinin/longphase), [whatshap](https://github.com/whatshap/whatshap) or [HiPhase](https://github.com/PacificBiosciences/HiPhase) | ||
- Create methylation pileups with [modkit](https://github.com/nanoporetech/modkit) | ||
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##### Annotation | ||
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- Annotate SNVs and INDELs with databases of choice, i.e. [gnomAD](https://gnomad.broadinstitute.org), [CADD](https://cadd.gs.washington.edu) etc. with [echtvar](https://github.com/brentp/echtvar) and [VEP](https://github.com/Ensembl/ensembl-vep) | ||
- Annotate repeat expansions with [stranger](https://github.com/Clinical-Genomics/stranger) | ||
- Annotate SVs with [SVDB](https://github.com/J35P312/SVDB) and [VEP](https://github.com/Ensembl/ensembl-vep) | ||
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##### Ranking | ||
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- Rank SNVs with [GENMOD](https://github.com/Clinical-Genomics/genmod) | ||
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## Usage | ||
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> [!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. | ||
Prepare a samplesheet with input data: | ||
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`samplesheet.csv` | ||
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``` | ||
project,sample,file,family_id,paternal_id,maternal_id,sex,phenotype | ||
NIST,HG002,/path/to/HG002.fastq.gz,FAM1,HG003,HG004,1,2 | ||
NIST,HG005,/path/to/HG005.bam,FAM1,HG003,HG004,2,1 | ||
``` | ||
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Supply a reference genome with `--fasta` and choose a matching `--preset` for your data (`revio`, `pacbio`, `ONT_R10`). Now, you can run the pipeline using: | ||
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```bash | ||
nextflow run genomic-medicine-sweden/nallo \ | ||
-profile <docker/singularity/.../institute> \ | ||
--input samplesheet.csv \ | ||
--preset <revio/pacbio/ONT_R10> \ | ||
--fasta <reference.fasta> \ | ||
--outdir <OUTDIR> | ||
``` | ||
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For more details and further functionality, please refer to the [usage documentation](https://github.com/genomic-medicine-sweden/nallo/blob/dev/docs/usage.md). | ||
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> [!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/usage/configuration#custom-configuration-files). | ||
To run in an offline environment, download the pipeline and singularity images using [`nf-core download`](https://nf-co.re/tools/#downloading-pipelines-for-offline-use): | ||
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``` | ||
nf-core download genomic-medicine-sweden/nallo | ||
``` | ||
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## Credits | ||
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genomic-medicine-sweden/nallo was originally written by Felix Lenner. | ||
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We thank the following people for their extensive assistance in the development of this pipeline: | ||
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## Contributions and Support | ||
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If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). | ||
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## Citations | ||
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If you use genomic-medicine-sweden/nallo for your analysis, please cite it using the following doi: [10.5281/zenodo.13748210](https://doi.org/10.5281/zenodo.13748210). | ||
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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/main/LICENSE). | ||
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> **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](https://dx.doi.org/10.1038/s41587-020-0439-x). | ||
An extensive list of references for the tools used by the pipeline can be found in the [`CITATIONS.md`](CITATIONS.md) file. |
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# based on: https://github.com/nf-cmgg/structural/blob/master/mkdocs.yml | ||
site_name: genomic-medicine-sweden/nallo | ||
repo_name: genomic-medicine-sweden/nallo | ||
repo_url: https://github.com/genomic-medicine-sweden/nallo | ||
site_url: https://genomic-medicine-sweden.github.io/nallo | ||
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nav: | ||
- Home: index.md | ||
- Usage: usage.md | ||
- Output: output.md | ||
- Parameters: parameters.md | ||
- Citations: CITATIONS.md | ||
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theme: | ||
name: material | ||
icon: | ||
logo: octicons/checklist-16 | ||
repo: fontawesome/brands/github | ||
palette: | ||
# Palette toggle for dark mode | ||
- media: "(prefers-color-scheme: dark)" | ||
scheme: slate | ||
primary: custom | ||
accent: custom | ||
toggle: | ||
icon: material/weather-sunny | ||
name: Switch to light mode | ||
# Palette toggle for light mode | ||
- media: "(prefers-color-scheme: light)" | ||
scheme: default | ||
primary: custom | ||
accent: custom | ||
toggle: | ||
icon: material/weather-night | ||
name: Switch to dark mode | ||
font: | ||
text: Inter | ||
features: | ||
- content.code.annotate | ||
- content.code.copy | ||
- navigation.instant | ||
- navigation.top | ||
- navigation.tracking | ||
- navigation.sections | ||
- navigation.footer | ||
- search.share | ||
- toc.follow | ||
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markdown_extensions: | ||
- attr_list | ||
- md_in_html | ||
- admonition | ||
- pymdownx.emoji: | ||
emoji_index: !!python/name:material.extensions.emoji.twemoji | ||
emoji_generator: !!python/name:material.extensions.emoji.to_svg | ||
- pymdownx.highlight: | ||
anchor_linenums: true | ||
line_spans: __span | ||
pygments_lang_class: true | ||
- pymdownx.inlinehilite | ||
- pymdownx.snippets | ||
- pymdownx.superfences | ||
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extra: | ||
version: | ||
provider: mike |