Look into freebayes. There is samtools/bcftools based option as well.
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sorted bam and bbmap
Is there an option for bbmap that will result in a sorted bam? I know adding "out=mapped.bam" this will produce a bam but it appears to be unsorted:
Thank youCode:samtools view -H /home/cmccabe/IDP/mapped_unmatched_adapter_removed_quality_trimmed_NA12878-NextSeq-S1_R1_and_R2.bam @HD VN:1.4 SO:unsorted
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Of course, I do my best to prevent needing people to do work, so...Originally posted by GenoMax View PostNo. You have to do some work
Code:$ samtools sort your_file.bam your_file_sorted
That will write a script "bs.sh" that contains the command to make a sorted, indexed bam file, then execute it after BBMap is done. It saves... well, at least a couple of keystrokesCode:bbmap.sh in=reads.fq out=mapped.bam bs=bs.sh; sh bs.sh
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Unfortunately, not easily, due to memory constraints. BBMap's shell script already grabs all available memory by default. It can launch a samtools process for conversion to bam, which is a low-memory streaming process; but sorting uses a lot of memory (to do it most efficiently), so it might run out of memory. Java cannot reduce its virtual memory allocation after starting, and won't share that memory with other programs.Originally posted by westerman View PostCouldn't you create the shell script and then execute it within BBMap?
It would work fine on most individual computers since physical memory limits would never be exceeded, but job schedulers often have strict limits on virtual memory and will kill jobs that exceed it. So, it would reduce stability (and typically fail) on clusters unless I did parsing in the shellscript to launch a new process after BBMap terminates, or the user launched BBMap with a custom -Xmx flag. I try to keep the shellscripts as simple as possible. Anyway, there are workarounds, but they all add complexity due to memory issues.
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by GATTACATLove this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
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by SEQadmin2
I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...-
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