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Old 04-12-2019, 01:45 AM   #1
jmc887
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Default Low yields with HyperPlus library kit

Hello everyone,

I am trying out the HyperPlus kit for library prep. The protocol says the expected pre-capture yield should be ≥ 1 ug. However, I got around 500 ng yield (my input was 100 ng). I have performed the protocol precisely, so I would like to know if anyone has more experience with this kit and if you have encountered the same issue?

Thanks in advance,
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Old 04-12-2019, 08:17 PM   #2
luc
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How many PCR cycles did you run? Do the libraries have the expected length?
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Old 04-14-2019, 11:31 PM   #3
jmc887
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I ran 7 cycles in total (as indicated for SeqCap adapters), and yes, the libraries have the expected length. Its only the yield that is the lowest I get, compared to other library prep kits using the same amount of input DNA (SureSelect and Nextera Flex).
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Old 04-15-2019, 07:04 AM   #4
jhalpin
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How did you measure your 100ng input?
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Old 04-19-2019, 12:47 AM   #5
jmc887
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Quote:
Originally Posted by jhalpin View Post
How did you measure your 100ng input?

I measured with Qubit
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Old 04-19-2019, 04:45 PM   #6
luc
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Sorry, I have no clue what could have gone wrong. We do see significantly higher yields with the same Kapa kit using Biooscientific adapters after 7 PCR cycles.
Did you run a two-sided size selection? This will significantly reduce yields.
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Old Yesterday, 11:25 PM   #7
jmc887
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Thanks for your answer luc, in fact yes, i did run a double-sided size selection. I will probably try again without this double-sided selection.
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