We are experiencing high enrichment with our emPCR as well as a high %Short when we achieve an enrichment that allows us to sequence (GS Junior). We have examined our PCR plates, we have used different thermal cyclers and we have tried different tissuelysers / turraxes (looking at emulsion formation) and nothing seems to provide any clues. The only parameter we haven't really played with are the temperatures, times and number of cycles the thermal cyclers of the program.
We are using the temps, times and cycles as outlined in the Roche protocol for em PCR (LVs, MVs, SVs and Junior kits). We are looking at 16S Amplicons. Has anyone else looked at this and use different temps, times and cycles?
We are using the temps, times and cycles as outlined in the Roche protocol for em PCR (LVs, MVs, SVs and Junior kits). We are looking at 16S Amplicons. Has anyone else looked at this and use different temps, times and cycles?
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