Hi all,
we are using Agilent SureSelect Target Enrichment System (protocol v1.5) and plan to work with ABI SOLiD.
We have problems as early in the protocol as target enrichment up to pre-hybridization amplification. After this amplification we analyzed our PCR product but there was no DNA amount detectable with NanoDrop. The starting DNA amounts were 9µg (whole genome amplification (WgA) done before target enrichment; preferred) or 6µg (without WgA; just in comparison to WgA sample).
Has anyone similar problems or suggestions what the problem could be?
Is there a possibility to check a successful amplification beside the PCR product itself? Something like a positive control?
Has anybody done an additional amplification step to increase the PCR product amount?
Hoping for help, thanks in advance,
Theresa
we are using Agilent SureSelect Target Enrichment System (protocol v1.5) and plan to work with ABI SOLiD.
We have problems as early in the protocol as target enrichment up to pre-hybridization amplification. After this amplification we analyzed our PCR product but there was no DNA amount detectable with NanoDrop. The starting DNA amounts were 9µg (whole genome amplification (WgA) done before target enrichment; preferred) or 6µg (without WgA; just in comparison to WgA sample).
Has anyone similar problems or suggestions what the problem could be?
Is there a possibility to check a successful amplification beside the PCR product itself? Something like a positive control?
Has anybody done an additional amplification step to increase the PCR product amount?
Hoping for help, thanks in advance,
Theresa
Comment