Comments (11)
At the moment I'm not sure that's possible (for what it's worth, PBAT was just coming out when this package was created). Can you point to any convenient (ideally smallish, but I'll take big and high quality over small and low quality) public PBAT datasets? I can then play around a bit and see what works reasonably (or add an option to MethylDackel if needed).
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Here is an example file where the image above comes from:
https://dl.dnanex.us/F/D/bygGZ2v3Y8qq8Pp8YGFyKyp81bp4xbJ67Bzk618X/CEG60-53-C1-oxBS_S1_L001_R1_001.1.cutB.seqtk_revcom.u.bwameth.paired.primary.barcoded.2Df.sorted.bam
https://dl.dnanex.us/F/D/kB19gVB0qjpP236Jzv9KjxBgjk7g7XgB0PqqQkPv/CEG60-53-C1-oxBS_S1_L001_R1_001.1.cutB.seqtk_revcom.u.bwameth.paired.primary.barcoded.2Df.sorted.bam.bai
It's small in size but has high coverage concentrated in certain regions.
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Wonderful, thanks!
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The reference is the standard GRCh38Decoy from NCBI:
https://dl.dnanex.us/F/D/jzGpFXgf85p8XQB3JVYY9fxYkFXVgQqP062Jx8v4/Homo_sapiens_NCBI_GRCh38Decoy.genome.fa
The reads where aligned with bwa-meth:
python bwa-meth/bwameth.py --threads 36 --reference include_indexes_dir/input/GRCh38Decoy.fa CEG60-53-C1-oxBS_S1_L001_R1_001.1.cutB.seqtk_revcom.u.fq.gz CEG60-53-C1-oxBS_S1_L001_R1_001.2.cutB.seqtk_revcom.u.fq.gz --prefix CEG60-53-C1-oxBS_S1_L001_R1_001.1.cutB.seqtk_revcom.u.bwameth
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Let me know if I can supply any more relevant details...
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Will do. I'm creating the indices at the moment.
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A small update, I have a modified version of bwameth with a --pbat
option that seems to be producing good results (I'm testing with DRR019425). I've only tested paired-end reads so far and need to give single-end reads a go. Once those are working nicely too I'll make a pull-request on bwameth, at which point you should be able to get some reasonable results with it. MethylDackel itself doesn't need any modifications, you can give it the BAM file from bismark (after sorting) and if bwameth with --pbat
is used the results should be pretty similar (I get correlation coefficients of ~90%, with bwameth producing a LOT more alignments in a fraction of the time).
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Please try the pbat
branch with the --pbat
option from my fork here. That now seems to be working correctly for SE and PE data.
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@avilella Did you have a chance to test that bwameth branch? If so, did it produce reasonable results?
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Related Issues (20)
- Understanding --minConversionEfficiency argument HOT 1
- Genome browsing from MethylDackel bedGraphCpG file
- per-fragment methylation HOT 2
- mbias result is different between bismark and bwameth output HOT 1
- CURL_OPENSSL conflict with samtools HOT 3
- Installation failure: "bigWig.h: No such file or directory" HOT 1
- Clarification on definition of "unmethylated C" HOT 1
- Coverage of C sites HOT 1
- mbias HOT 1
- Does indel effect the methylation calling or C context determination HOT 1
- Positions in cytosine_report did not match the regions in providing bed file
- about M-bias HOT 1
- Could not repeat a CpG extraction with the same reference file and its index
- Mixed up reads within bam file
- How to index genome file for MethylDackel? HOT 1
- Confused regarding CTOT, CTOB. Are there suggested values? HOT 3
- Issue running MethylDackel extract in parallel mode using minConversionEfficiency
- Question about minimum coverage
- Different CpG calls when using different regions of inclusion for Methyldackel extract
- Alignment trimming for Soft Clip reads?
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