Daniel:Notebook/ComboLock/2017-4-3

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Revision as of 19:51, 3 April 2017 by >Djacobse (Created page with "=RCA Test (Started March 24)= Back to Calendar I'll try qPCR on the RCA product again, this time on th...")
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RCA Test (Started March 24)

Back to Calendar


I'll try qPCR on the RCA product again, this time on the NEB 12 hr samples (since I am running low on X samples), but for few cycles and with only the forward primer (which should be RC to the RCA product. This will be fed into Bglii digestion

  1. qPCR-Second strand synthesis
    1. Make the following 13.2X master mix
      1. 13.2 uL AmpF-6.4Sol (10 uM)
      2. 264 uL Kapa SYBR 2X Master Mix
      3. 224.4 uL nfH2O
    2. Aliquot 38 uL master mix into each lane
    3. Add 2 uL sample according to plate layout
    4. File:PlateLayout-20170403-RCATest-CPLP-ssSynthesis.png
    5. Run the following protocol
      1. 95C for 2 min
      2. 95C for 5 sec
      3. 55C for 30 sec
      4. 72C for 45 sec
      5. Goto (b) x20
      6. 72C for 2 min
  2. Qiaquick Column
  3. TBE Gel
  4. Bgl Digestion