Dear community,
maybe someone of you has an answer.
We perform a Hybrid Capture Library Enrichment protocol. In most cases the concentration and sequence coverage of the final library is in the expected range. In some cases, however, we get high concentrations but very low genomic coverage, indicative for ineffective hybridization.
I assumed that I might have misquantified the libraries before pooling (or errrors during pooling) and thus have to much DNA in the pools. I tried to counter it by slightly increasing (by 1µl) the cotDNA Volume I add to each pool before hybridization. I assumed that that this might increase the hybridzation efficiency (or at least would not harm it). However, the opposit was the case. The test pools all had a severely decreased concentration (about half).
Finally the question:
1. What might be the reason for this effect?
2. Would a decrease in cotDNA, instead, might help?
Thanks for your help.
Andreas
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