CRAM to BAM conversion

This workflow is used in IGSR for:

  1. Downloading a CRAM file from the archive (ENA, IGSR FTP, etc…)
  2. Convert it to BAM
  3. Create an index for converted BAM.

This workflow relies on the ASPERA service for the fast download of the data from the archives

Dependencies

  • Nextflow

This pipeline uses a workflow management system named Nextflow. This software can be downloaded from:

https://www.nextflow.io/

  • SAMTools
  • Aspera connect software:

How to run the pipeline

  • First, you need to create a nexflow.config file that can be used by Nextflow to set the required variables. Here goes an example of one of these files:

    params.samtools_folder='~/bin/samtools-1.9/' // folder containin the samtools binary
// params defaults for ascp client
params.key_file = '/homes/ernesto/.aspera/connect/etc/asperaweb_id_dsa.openssh' // Private-key file name (id_rsa) for authentication
params.transfer_rate = '900M'
params.port = 33001 // TCP port used for SSH authentication

  • Then, you can start your pipeline by doing:

    nextflow -C nextflow.config run $IGSR_CODEBASE/scripts/FILE/cram2bam.nf --file input.txt
Where:

  • -C option allows you to specify the path to the nextflow.config file

  • $IGSR_CODEBASE is the folder containing the igsr codebase downloaded from https://github.com/igsr/igsr_analysis.git

  • --file File with the urls pointing to the CRAM files to be converted. This file shold have a content similar to:

    url,dest,prefix
era-fasp@fasp.sra.ebi.ac.uk:/vol1/ERZ454/ERZ454001/ERR1457180.cram,/path/in/dest/ERR1457180.cram,ERR1457180

    Where url points to the location of the file to be downloaded, dest is the path in the local machine where it will be downloaded and prefix is used as the string used in the converted BAM file and its respective index

Pipeline output

This worklow will create a folder name converted/ with 2 output files:

  • prefix.bam
BAM file resulting after converting the downloaded CRAM file
  • prefix.bam.bai
The index created after running samtools index prefix.bam