Illumina library

[Katerina Cvikova]

Hi,
do you have experience with good results of illumina libraries from Agilent, but qPCR says that there are not anything? I prepared library (DNA, TruSeq), measured it with qPCR and Agilent, everything was OK and friezed it. Before sequencing (1week) I did Agilent and qPCR again and according to qPCR there were not anything. Is is possible that DNA de-gradated so fast, or is it some contamination that damage DNA or something else? I did a new library and new library with new kit and qPCR KO, agilent OK.
Thanks for advice.

[nucacidhunter]

Fast library degradation is unlikely. If Agilent (I assume BA or TapeStation) indicates DNA presence, the issue could be qPCR.

[Katerina Cvikova]

Yes, I assumed this version too. But I took a new kit and prepare new buffer for qPCR an nothing. Then I prepared a new libtary with Neb kit and results from Agilent and qPCR are OK. I had a similar problem with Nextera 2 years ago and there were really nothing, I tried to sequenced it.