Introduction

sanger-tol/genomenote is a bioinformatics pipeline that takes aligned HiC reads, creates contact maps and chromosomal grid using Cooler, and display on a HiGlass server. The pipeline also collates (1) assembly information, statistics and chromosome details from NCBI datasets, (2) genome completeness from BUSCO, (3) consensus quality and k-mer completeness from MerquryFK, and (4) HiC primary mapped percentage from samtools flagstat.

1. Fetches genome metadata from ENA, NCBI, and GoaT
2. Summary statistics (NCBI datasets summary genome accession)
3. Convert alignment to BED (samtools view, bedtools bamtobed)
4. Filter BED (GNU sort, filter bed)
5. Contact maps (Cooler cload, Cooler zoomify, Cooler dump)
6. Genome completeness (NCBI API, BUSCO)
7. Consensus quality and k-mer completeness (FASTK, MERQURY.FK)
8. Collated summary table (createtable)
9. Present results and visualisations (MultiQC, R)

Usage

Note If you are new to Nextflow and nf-core, please refer to this page on how to set-up Nextflow. Make sure to test your setup with -profile test before running the workflow on actual data.

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

samplesheet.csv:

sample,datatype,datafile
mMelMel3,hic,/analysis/mMelMel3.2_paternal_haplotype/read_mapping/hic/GCA_922984935.2.unmasked.hic.mMelMel3.cram
mMelMel3,pacbio,/genomic_data/mMelMel3/pacbio/kmer/k31

Each row represents an aligned HiC reads file, an unaligned PacBio/10X reads file, or a PacBio/10X k-mer database.

Now, you can run the pipeline using:

nextflow run sanger-tol/genomenote \
   -profile <docker/singularity/.../institute> \
   --input samplesheet.csv \
   --fasta genome.fasta \
   --assembly GCA_922984935.2 \
   --bioproject PRJEB49353 \
   --biosample SAMEA7524400 \
   --outdir <OUTDIR>

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.

For more details, please refer to the usage documentation and the parameter documentation.

Credits

sanger-tol/genomenote was originally written by Priyanka Surana.

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

• Matthieu Muffato
• Beth Yates
• Shane McCarthy and Yumi Sims for providing software and algorithm guidance.
• Cibin Sadasivan Baby for providing reviews.

Contributions and Support

If you would like to contribute to this pipeline, please see the contributing guidelines.

Citations

If you use sanger-tol/genomenote for your analysis, please cite it using the following doi: 10.5281/zenodo.7949384

An extensive list of references for the tools used by the pipeline can be found in the CITATIONS.md file.

This pipeline uses code and infrastructure developed and maintained by the nf-core community, reused here under the MIT license.

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.