Version 1

Workflow Type: Nextflow


A fully reproducible ancient and modern DNA pipeline in Nextflow and with cloud support..

Build Status Nextflow Slack Statusinstall with bioconda Docker Container available Singularity Container available DOI


nf-core/eager is a bioinformatics best-practice analysis pipeline for NGS sequencing based ancient DNA (aDNA) data analysis.

The pipeline uses Nextflow, a bioinformatics workflow tool. It pre-processes raw data from FASTQ inputs, aligns the reads and performs extensive general NGS and aDNA specific quality-control on the results. It comes with docker, singularity or conda containers making installation trivial and results highly reproducible.

Pipeline steps

By default the pipeline currently performs the following:

  • Create reference genome indices for mapping (bwa, samtools, and picard)
  • Sequencing quality control (FastQC)
  • Sequencing adapter removal and for paired end data merging (AdapterRemoval)
  • Read mapping to reference using (bwa aln, bwa mem or CircularMapper)
  • Post-mapping processing, statistics and conversion to bam (samtools)
  • Ancient DNA C-to-T damage pattern visualisation (DamageProfiler)
  • PCR duplicate removal (DeDup or MarkDuplicates)
  • Post-mapping statistics and BAM quality control (Qualimap)
  • Library Complexity Estimation (preseq)
  • Overall pipeline statistics summaries (MultiQC)

Additional functionality contained by the pipeline currently includes:

  • Illumina two-coloured sequencer poly-G tail removal (fastp)
  • Automatic conversion of unmapped reads to FASTQ (samtools)
  • Damage removal/clipping for UDG+/UDG-half treatment protocols (BamUtil)
  • Damage reads extraction and assessment (PMDTools)

Quick Start

  1. Install nextflow

  2. Install one of docker, singularity or conda

  3. Download the EAGER pipeline

bash nextflow pull nf-core/eager

  1. Test the pipeline using the provided test data

bash nextflow run nf-core/eager -profile <docker/singularity/conda>,test --pairedEnd

  1. Start running your own ancient DNA analysis!

bash nextflow run nf-core/eager -profile <docker/singularity/conda> --reads'*_R{1,2}.fastq.gz' --fasta '<REFERENCE>.fasta'

NB. You can see an overview of the run in the MultiQC report located at <OUTPUT_DIR>/MultiQC/multiqc_report.html

Modifications to the default pipeline are easily made using various options as described in the documentation.


The nf-core/eager pipeline comes with documentation about the pipeline, found in the docs/ directory or on the main homepage of the nf-core project:

  1. Nextflow Installation
  2. Pipeline configuration
  3. Running the pipeline
  4. Output and how to interpret the results
  5. Troubleshooting
  6. EAGER Troubleshooting


This pipeline was written by Alexander Peltzer (apeltzer), with major contributions from Stephen Clayton, ideas and documentation from James A. Fellows Yates, Raphael Eisenhofer, Maxime Borry and Judith Neukamm. If you want to contribute, please open an issue and ask to be added to the project - happy to do so and everyone is welcome to contribute here!


If you've contributed and you're missing in here, please let me know and I'll add you in.

Tool References

Total size: 216 KB
help Creators and Submitter
  • Alexander Peltzer
  • Stephen Clayton
  • James A. Fellows Yates
  • Maxime Borry

Views: 980   Downloads: 23

Created: 25th Feb 2020 at 11:05

Last updated: 25th Feb 2020 at 15:19

Last used: 27th Jan 2022 at 06:37

help Attributions


Version History

Version 1 (earliest) Created 25th Feb 2020 at 11:05 by Finn Bacall

Added/updated 33 files

Open master 0137199

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