Daniel:Notebook/ComboLock/2017-3-29

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Revision as of 17:41, 29 March 2017 by >Djacobse (→‎Bglii Digestion)
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RCA Test (Started March 24)

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Bglii Digestion

Testing bglii digestion of RCA product. No qPCR to start with (RCA should be enough)

  1. Bglii digestion
    1. Make the following reactions; Do not add Bglii yet
    2. Reagent Single Rxn Vol Master Mix (30.2X)
      RCA Rxn 2 0
      Bglii Cutter Primer + (100 uM) 2 60.4
      10X Buffer 3.1 2 60.4
      Bglii 1 0
      nfH2O 13 392.6
      Total 20 513.4
    3. Use the following thermocycler program
      1. 5 min 95C
      2. Ramp to 50C at 0.2C/s
      3. 10 min 50C
      4. 2 hr 37C; When this cycle starts add 1 uL Bglii
      5. Heat kill with 65C for 20 min