Hello,
We have had library prep problems using the neoprep for the last 6 runs in a row. It seems like the neoprep library output is failing and/or generating heavily 3' biased libraries with low diversity despite loading 100ng (spec range is 25-100ng) of RNA with RINs of 10, control UHR samples, and samples that previously worked and undergoing 15cycles of PCR.
We have re-bioanalyzed samples after these failures and they always checked out. We changed all our plastic ware and changed lots of reagents.
We have run over 40 neoprep runs without very many problems with the exception of last fall when we had a grouping of similar failed runs. So we thought that perhaps the change in temperature was causing dry indoor air and static. So we humidified the air and grounded all our consumables to dissipate static. Still no luck.
So in general library chemistry, if starting with good input and quality RNA does anyone have any idea how we can be losing so much diversity and gaining strong 3'bias?
We have had library prep problems using the neoprep for the last 6 runs in a row. It seems like the neoprep library output is failing and/or generating heavily 3' biased libraries with low diversity despite loading 100ng (spec range is 25-100ng) of RNA with RINs of 10, control UHR samples, and samples that previously worked and undergoing 15cycles of PCR.
We have re-bioanalyzed samples after these failures and they always checked out. We changed all our plastic ware and changed lots of reagents.
We have run over 40 neoprep runs without very many problems with the exception of last fall when we had a grouping of similar failed runs. So we thought that perhaps the change in temperature was causing dry indoor air and static. So we humidified the air and grounded all our consumables to dissipate static. Still no luck.
So in general library chemistry, if starting with good input and quality RNA does anyone have any idea how we can be losing so much diversity and gaining strong 3'bias?
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