Daniel:Notebook/ComboLock/2016-8-29

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Revision as of 16:33, 29 August 2016 by >Djacobse (Created page with "=Positive Control Dilution Test (Started Friday)= Back to Calendar ==Protocol== Today I'm testing the ...")
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Positive Control Dilution Test (Started Friday)

Back to Calendar

Protocol

Today I'm testing the samples using the P2 and P12RC amplification primers. Yesterday included the original amplicon materials; this qPCR should be only the DNA still attached to beads.

  1. qPCR X2-Primers
    1. Set up 14 reaction with 2 uL sample, 25 uL KAPA, 22 uL nfH2O, 0.5 uL primer2 and 0.5 uL primer12RC
    2. Master Mix
      1. 352.5 uL KAPA SyberFast MM
      2. 310.2 uL nfH2O
      3. 7.05 uL primer2
      4. 7.05 uL primer12RC
    3. Add 48 uL Master Mix to each well
    4. qPCR Cycles
      1. 95C 3 min
      2. 95C 3 sec
      3. 55C 30 sec
      4. 72C 20 sec
      5. plate read
      6. goto b x32
      7. 72C 2 min
      8. 16C hold