<div dir="ltr">Hi Titus and the khmer list,<div>I'm working on transcriptome assembly with samples treated at 12 different temperatures to capture genes expressed across the thermal range of my favorite ant species. I pooled the samples and ran them in a single lane of 100 bp paired end HiSeq, so I have about 16 million reads per sample, 160 million reads total. </div>
<div><br></div><div>My question: </div><div>is there any benefit to merging my paired-end reads (e.g. using FLASH <a href="http://bioinformatics.oxfordjournals.org/content/early/2011/09/07/bioinformatics.btr507" target="_blank">http://bioinformatics.oxfordjournals.org/content/early/2011/09/07/bioinformatics.btr507</a>) prior to running diginorm? A preliminary run of FLASH on some of my samples showed that about 65% of reads are merged (which is a bit surprising since the library was supposed to have been size-selected at 200 bp). </div>
<div><div><br></div><div>My thought is to run diginorm on the merged reads, and also on the un-merged reads using the `-p` option as documented previously (<a href="http://lists.idyll.org/pipermail/khmer/2013-July/000123.html">http://lists.idyll.org/pipermail/khmer/2013-July/000123.html</a>). I'd then combine all these and run a second pass of diginorm.</div>
<div><br></div><div>Is this a valid approach, or is merging reads redundant with what diginorm does (since reads that add extra coverage would be tossed out anyway)? </div><div><br></div><div>Apologies if this is a noob question.</div>
<div><br></div><div>Thanks for the software!</div><div><br></div><div>-John</div><div><br></div>-- <br><div dir="ltr"><div><span style="color:rgb(102,102,102)">Postdoctoral Research Associate</span><br></div><div><span style><font color="#666666">Department of Biology, University of Vermont</font></span></div>
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