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Old 01-25-2013, 08:59 AM   #1
dummyseq
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Default low DNA conc after amplification -DNA library prep

I am starting with 500ng DNA [using cell lines] for DNA library prep using SureSelect XT protocol. I am getting very low DNA conc after amplification ,using 10 cycles. It is not enough for me to hybridise for Hiseq.

What could be inhibiting my DNA conc....changed all reagents,using brand new kit ....same kit being used by others in the lab and is working...

Any advice appreciated.
Thanks
Dummyseq
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Old 01-25-2013, 10:56 AM   #2
kwaraska
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What quantification method are you using? They suggest nanodrop which gives higher readings than BioA.

When I did it and used my BioAnalyzer results I did not have enough but when characterized by nanodrop, I was fine. I think they use as high a number as they do assuming a nanodrop reading that may be wildly off.
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Old 01-25-2013, 11:26 AM   #3
dummyseq
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I am using TapeStation ...
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