[lilyhu]

We also suffered from low enrichment problem, mostly below 2%. The template that we are using is amplicon with fusion primer (with adaptor and barcode). We have checked the template with bioanalyzer and KAPA qPCR first. Then we used 1.7 cpb in the LV Lib-L emPCR. The enrichment is lower than 2% that we suspected the cpb not optimize, emPCR failed or the problem of the template? Then we went for SV using various cpb from 0.25 to 17, again the enrichment was <2% while the positive control (from previous work fine library) is OK but cpb increased from 1.7 to 14.6.

I am working on library instead of amplicon. I thought cpb is a relative number depending how you test you library concentration.

[boerm]

preincubation of enrichment beads with enrichment primer

So, did it help? We are still doing it. Current work-around is to underload the plate and simply expect to have to do two runs to get the number of reads that we routinely got up until Dec 2011. Been nearly a full year of this, now.

Dear HMorrison,

Yes, preincubation of the enrichment beads with the enrichment primer as you suggested worked in our LV lib-L. The problem now is that the enrichments is now suddenly 40%, while from the SV (done without preincubation step) with the same cpb (1,5) gave 5% enrichment.