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Old 09-27-2017, 07:01 AM   #1
RickC7
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Location: Baton Rouge, Louisiana

Join Date: Feb 2010
Posts: 30
Default Index Read distribution

Hello,

I'm looking for advice, help, thoughts, etc on how to achieve more balanced read output when multiplexing. Typically I run all libraries on Agilent Bioanalyzer and pool according to the library peak in equimolar amounts. My last TruSeq mRNA seq project had between 2% and 8% reads for each sample, which varied read output 12 to 48million reads per sample. I see variations also when doing 16s(Schloss barcodes). I just wondering if variation is typical, some barcodes may amplify better than others, etc?? Do most labs see more even read distribution or is it more likely to "do the best you can and hope the outcome is acceptable"?

I'd be interested in any success stories using normalization protocols, plates, beads etc for library pooling.

Thanks for any help!

Last edited by RickC7; 09-27-2017 at 08:06 AM.
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