How to perform gradient PCR?

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    • #18016
      w19910411
      Participant

      How should one perform gradient PCR, and how much time will it take?

    • #115795
      Javedz2001
      Participant

      The time will remain the same. Set up PCR as follows:

      FAST Cycling Conditions Temp (°C) Time (sec) Initial denaturation/Hot Start 95 30 Steps 1–3 are repeated through 40 cycles
      Step 1: 95 for 10 seconds
      Step 2: 54–70 (gradient) for 15 seconds
      Step 3: 72 15-60 seconds (depends on the length of your amplicon)

      Good Luck: Javed Siddiqi

    • #116000
      Fornita
      Participant
      quote Javedz2001:

      The time will remain the same. Set up PCR as follows:
      Step 2: 54–70 (gradient) for 15 seconds
      Good Luck: Javed Siddiqi

      For step 2, you could change the temp. gradient according to your sequences properties. lower of higher.

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