• Kingfisher
  • Installation
  • Usage
    • Method details
    • Near parity with ena-fast-download
  • FAQ
  • License

Kingfisher

Kingfisher is a fast and flexible program for procurement of sequence files from public data sources, including the European Nucleotide Archive (ENA), NCBI SRA, Amazon AWS and Google Cloud. It's input is an "Run" accession e.g. DRR001970.

It attempts to download data from a series of methods, which it attempts in order until one works. Then the downloaded data is converted to an output SRA / FASTQ / FASTA / GZIP file format as required. Both download and extraction phases are usually quicker than using the NCBI's SRA toolkit.

This software was originally known as ena-fast-download. Kingfisher implements almost all of that tool's functionality, but also handles data sources other than ENA. See the 'Usage' section for the equivalent invocation.

The ena-ascp method of this tool was built based on the very helpful bio-stars thread https://www.biostars.org/p/325010/ written by @ATpoint. To find run identifiers to be used as input to kingfisher, you might find the SRA explorer site helpful.

Installation

Kingfisher can be installed by installing its conda dependencies as follows

conda create -c conda-forge -c bioconda -n kingfisher pigz python extern curl sra-tools
conda activate kingfisher
git clone https://github.com/wwood/kingfisher-download
cd kingfisher-download/bin
export PATH=$PWD:$PATH
kingfisher -h

Optionally, to use the ena-ascp method, an Aspera connect client is also required. See https://www.ibm.com/aspera/connect/ or https://www.biostars.org/p/325010/

Usage

kingfisher get -r ERR1739691 -m ena-ascp aws-http prefetch

This will download .fastq.gz files of the run ERR1739691 from the ENA, or failing that, downloads an .sra file from the Amazon AWA Open Data Program and then converts to FASTQ, or failing that use NCBI prefetch to download and convert that to FASTQ.

Output files are put into the current working directory.

Method details

Near parity with ena-fast-download

Ena-fast-download was the original name for this tool. To imitate that tool's functionality:

kingfisher get -r ERR1739691 -m ena-ascp

FAQ

If you see this error /bin/sh: 1: ascp: not found as below:

$ kingfisher get -r ERR3357550 -m ena-ascpkingfisher -r ERR3357550 -m ena-ascp
05/04/2021 05:13:45 AM INFO: Attempting download method ena-ascp ..
05/04/2021 05:13:45 AM INFO: Using aspera ssh key file: $HOME/.aspera/connect/etc/asperaweb_id_dsa.openssh
05/04/2021 05:13:45 AM INFO: Querying ENA for FTP paths for ERR3357550..
05/04/2021 05:13:49 AM INFO: Downloading 2 FTP read set(s): ftp.sra.ebi.ac.uk/vol1/fastq/ERR335/000/ERR3357550/ERR3357550_1.fastq.gz, ftp.sra.ebi.ac.uk/vol1/fastq/ERR335/000/ERR3357550/ERR3357550_2.fastq.gz
05/04/2021 05:13:49 AM INFO: Running command: ascp -T -l 300m -P33001  -i $HOME/.aspera/connect/etc/asperaweb_id_dsa.openssh era-fasp@fasp.sra.ebi.ac.uk:/vol1/fastq/ERR335/000/ERR3357550/ERR3357550_1.fastq.gz .
/bin/sh: 1: ascp: not found
05/04/2021 05:13:49 AM WARNING: Error downloading from ENA with ASCP: Command 'ascp -T -l 300m -P33001  -i $HOME/.aspera/connect/etc/asperaweb_id_dsa.openssh era-fasp@fasp.sra.ebi.ac.uk:/vol1/fastq/ERR335/000/ERR3357550/ERR3357550_1.fastq.gz .' returned non-zero exit status 127.
05/04/2021 05:13:49 AM WARNING: Method ena-ascp failed
Traceback (most recent call last):
  File "./bin/kingfisher", line 330, in <module>
    raise Exception("No more specified download methods, cannot continue")
Exception: No more specified download methods, cannot continue

then you have not installed the Aspera client correctly. See the Installation section of this document.

License

Copyright Ben Woodcroft 2019-2021. Licensed under GPL3+. See LICENSE.txt.