nf-core / nf-core/chipseq

MACS3 is reporting that 100 paired reads are needed, even when single end data is used as input

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bug
Dominant language
Nextflow
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Description

Description of the bug

I have some low quality data where the second paired read had very low quality. As such, I tried to run chipseq version 2.1.0 with single-ended data as input.

This results in an error with the MACS3 step. MACS3 is reporting that 100 paired reads are needed:

WARNING @ 09 Jul 2026 02:09:00: [86 MB] #2 MACS3 needs at least 100 paired peaks at + and - strand to build the model, but can only find 0! Please make your MFOLD range broader and try again. If MACS3 still can't build the model, we suggest to use --nomodel and --extsize 147 or other fixed number instead.

Command used and terminal output
nextflow run nf-core/chipseq --input samplesheet.csv --read_length 150 --outdir mouse_output_single_end2 --genome GRCm38 -profile singularity -r 2.1.0
Relevant files

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System information

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First steps

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Research direction

Reproduce the report with the provided nf-core/chipseq command and single-end samples, then inspect the MACS3 step's handling of paired versus single-end input. Done means a single-end run no longer reports that paired reads are required and the MACS3 step proceeds successfully.

Written by the indexing model from the issue text.

Assessment

Domain
bioinformatics
Issue type
Bug
Difficulty
3/5
Estimated time
1-2 days
Activity status
Quiet
Clarity
Mostly clear
Newbie friendliness
48/100

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