So if you start with pure genomic DNA and do a TruSeq DNA kit reaction on it. Prior to enrichment PCR what % of the molecules will be replication-competent?
We just started doing qPCRs on all our libraries prior to enrichment PCR. I have not done an exhaustive study of the results, but I think we get results consistent with 10-30% of the molecules being "amplicons", as assayed using qPCR. By this I mean, you calculate your sample concentration by mass (using a BioAnalyzer DNA chip) and compare this to the concentration determined using a Kapa qPCR kit.
Does anyone else check this? If so, what do you see?
--
Phillip
We just started doing qPCRs on all our libraries prior to enrichment PCR. I have not done an exhaustive study of the results, but I think we get results consistent with 10-30% of the molecules being "amplicons", as assayed using qPCR. By this I mean, you calculate your sample concentration by mass (using a BioAnalyzer DNA chip) and compare this to the concentration determined using a Kapa qPCR kit.
Does anyone else check this? If so, what do you see?
--
Phillip
Comment