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-   -   converting bam files to non-normalized read counts (http://seqanswers.com/forums/showthread.php?t=11263)

lpn 05-10-2011 10:31 AM

converting bam files to non-normalized read counts
 
I am new to this business, so I would like to apologize in advance if this is trivial.

I have bam files produced to Tophat and need to run a differential gene expression analysis. I can do that with Cufflinks, but I also would like to compare the results with e.g. edgeR. However edgeR uses the raw counts, not normalized FPKMs. I can naively convert FPKMs to counts by multiplying to the gene length and total reads, but is there a better way to convert bam files to non-normalized read counts?

kopi-o 05-10-2011 12:45 PM

Check out the following:

http://www-huber.embl.de/users/ander...doc/count.html

http://sandberg.cmb.ki.se/media/data...mforgenes.html

http://code.google.com/p/bedtools/

2004rs 05-10-2011 02:23 PM

how to do this in Galaxy?
 
Another newbie question - is this capability integrated into Galaxy? I am having trouble figuring out the Galaxy "Operating on Genomic Intervals" tools capabilities. I have sequencing reads mapped using bowtie and would like to find the read count for reads that fall into genomic intervals tables downloaded from the UCSC Table Browswer, like sno/miRNAs or RefSeq genes.
Alternatively I can go outside Galaxy and use some of the tools that kopi-o suggested.
Thanks!

wangxj 10-09-2012 05:50 PM

Hello,
I have a question: how can you count the number of reads that mapped to each transcript? Now i have the bam file from the Tophat.

thanks

fubar 10-09-2012 07:52 PM

Quote:

Originally Posted by 2004rs (Post 41328)
Another newbie question - is this capability integrated into Galaxy?

The answer depends on exactly what you mean by "into Galaxy" :)
1. No, not on the public Galaxy instances run by the Galaxy Team.
2. Yes, if you have your own Galaxy. There's at least one suitable tool I seem to remember seeing in the main Toolshed - but it required Matlab (!) for some strange reason... OTOH, you can find a version that uses pysam (warning: work in progress!) at https://bitbucket.org/fubar/rgalaxy/...ics/bams2mx.py and https://bitbucket.org/fubar/rgalaxy/...cs/bams2mx.xml if you're game to try it. There's a bunch of other downstream tools in that repo that can use the resulting count matrixes including edgeR, DESeq and further downstream, GSEA and SPIA wrappers too. Enjoy but remember, YMMV.


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