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Discrepancy between bigwigs generated by bedtools and deeptools #123

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Caffeinated-Code opened this issue Oct 19, 2023 · 0 comments
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Caffeinated-Code commented Oct 19, 2023

Description of the bug

Two sets of bigWig coverage graphs are generated using bedtools and deeptools in v2.1.1.
The deeptools bigWig looks accurate and captures the bidirectional strand-specific signatures (green track).
But, the bedtools bigWigs on both strands are almost mirror images (brown track) - strand-specificity is lost (brown track).

Deeptools addresses and handles this strand-specificity issue:
Deeptools BamCoverage
Biostars Discussion

UCSC Browser/IGV visualization of these bidirectional transcriptional signatures from deeptools is helpful in guessing transcriptional start sites that can also be confirmed with PINTS/dREG peak calls.
In order to avoid confusion, future pipeline versions can drop generating bedtools coverage graphs.

Command used and terminal output

# Bedtools command run by nascent pipeline
bedtools genomecov -ibam sample.umi_dedup.sorted.bam -bg -strand + > sample.plus.bedGraph

# Deeptools command run by nascent pipeline
bamCoverage --bam sample.umi_dedup.sorted.bam --filterRNAstrand forward --numberOfProcessors 2 --outFileName sample.plus.bigWig

# Deeptools also generates bedgraphs if needed with this option
--outFileFormat bedgraph

Relevant files

bedtoolsVsdeeptools

System information

Nextflow Version: 23.04.1
Hardware: AWS Cloud9
Executor: Local
Container engine: Docker
OS: cpe:2.3:o:amazon:amazon_linux:2
Nascent Pipeline Version: v2.1.1

@Caffeinated-Code Caffeinated-Code added the bug Something isn't working label Oct 19, 2023
@edmundmiller edmundmiller self-assigned this Oct 19, 2023
@edmundmiller edmundmiller added this to the v2.2.0 milestone Oct 19, 2023
@edmundmiller edmundmiller modified the milestones: v2.2.0, 2.3.0 Feb 14, 2024
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