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  • 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,

  • #2
    How many PCR cycles did you run? Do the libraries have the expected length?

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    • #3
      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).
      Attached Files

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      • #4
        How did you measure your 100ng input?

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        • #5
          Originally posted by jhalpin View Post
          How did you measure your 100ng input?

          I measured with Qubit

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          • #6
            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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            • #7
              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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