nested pcr

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    • #11949
      shuk08
      Participant

      How to get rid of larger non-specific bands appearing in 1st round of pcr just above the expected bands (of approx. 1.5 kb)? The non-specific bands from the 1st pcr are amplified in 2nd round of pcr using nested pair of primer.

    • #93380
      JackBean
      Participant

      Are you asking or just saying us?

    • #93384
      canalon
      Participant

      I would try to optimize the first rund of PCR. Tips and tricks to do thta can be found here:
      http://info.med.yale.edu/genetics/ward/tavi/Guide.html

    • #93505
      roniadam
      Participant

      I had the same problem before. I advice you to use an adjuvant to improve your PCR effeciency. I used Q solution that comes with hotstar PCR master mix and it gives me a very good amplicon.

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