Hello everyone!
I have a question, I´m using NEB ultra prep DNA for my amplicom libraries (total size 150+120).
In the last step, when finish the PCR for the enrichment of adaptor ligated DNA I chek it in agarosa gel and I see a smear with a band in 600-1000 pb. This band is diferent in each library, some library 700, anothe 800...
I run a qPCR for quantification the librarys and I obtained very mixed result, from 1,5 nM to 1600nM...
The last time I sequenced the same library i obteined underclustering. I run in miniseq and it was 33k/mm2 and I think that it is the problem.
somebody could help me??
Thank you
I have a question, I´m using NEB ultra prep DNA for my amplicom libraries (total size 150+120).
In the last step, when finish the PCR for the enrichment of adaptor ligated DNA I chek it in agarosa gel and I see a smear with a band in 600-1000 pb. This band is diferent in each library, some library 700, anothe 800...
I run a qPCR for quantification the librarys and I obtained very mixed result, from 1,5 nM to 1600nM...
The last time I sequenced the same library i obteined underclustering. I run in miniseq and it was 33k/mm2 and I think that it is the problem.
somebody could help me??
Thank you
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