wellington_bootstrap questions

[jean997]

Hi there, This is probably a very simple question but I wanted to make sure that I was correctly understanding how to use the wellington_bootstrap.py script.

The script appears to only take two bam files "treatment_bam" and "control_bam". Is the expectation that if I have more than one sample in each group I will merge the bam files? It would be great if there was a way to pass multiple files for each group since the merged files can get very large! Thanks! Jean

[jpiper]

Ah, I might be able to modify the scripts to take arrays of BAM files, so that you can provide multiple treatments and controls.

I've noticed that someone has made a branch that does this here - https://github.com/PanosFirmpas/pyDNase/ for wellington_footprints.py, it hopefully shouldn't be too complicated to add support for wellington_bootstrap on top of this.

In the meantime, yes, you'd need to merge the BAM files. You can do this quickly using the samtools cat command provided all the BAM files share the same identical sequence dictionary

samtools cat [-h header.sam] [-o out.bam] <in1.bam> <in2.bam> [ ... ]

To generate the header.sam you can just use samtools view -H <in1.bam> > header.sam

[jean997]

Thanks - that's great! I was also wondering a few more things

1. Is there a way to interpret Wellington bootstrap scores in terms of p-values or false discovery rates? How do you choose a score cutoff. I know in the paper a score of 10 was chosen but I'm not exactly sure how this threshold was arrived at.
2. In the output of the footprint files I notice that there is a column (the 6th) that is entirely '+' symbols. Does this column mean something? Thanks! Jean

On Thu, Jun 30, 2016 at 8:42 PM, Jason Piper [email protected] wrote:

Ah, I might be able to modify the scripts to take arrays of BAM files, so that you can provide multiple treatments and controls.

I've noticed that someone has made a branch that does this here - https://github.com/PanosFirmpas/pyDNase/ for wellington_footprints.py, it hopefully shouldn't be too complicated to add support for wellington_bootstrap on top of this.

In the meantime, yes, you'd need to merge the BAM files. You can do this quickly using the samtools cat command provided all the BAM files share the same identical sequence dictionary

samtools cat [-h header.sam] [-o out.bam] <in1.bam> <in2.bam> [ ... ]

To generate the header.sam you can just use samtools view -H <in1.bam> > header.sam

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[jpiper]

1. Eurgh, it's been so long since I wrote that paper I need to have get into the right headspace and remember how the algorithm works, ha!

2. The is part of the BED specification, you can ignore these!