Daniel:Notebook/ComboLock/2017-4-7

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RCA Test CPLP (Started March 24)

Back to Calendar

Bgl Digestion-Minus Strand

  1. Bglii Digestion
    1. Make the following reactions
    2. Reagent Single Rxn Vol Master Mix + (4.2X) Master Mix - (4.2X)
      RCA Rxn 4 0 0
      RCA Primer + (10 uM) 2 8.4 0
      RCA Primer + (10 uM) 2 0 8.4
      10X Buffer 3.1 2 8.4 8.4
      Bglii 1 0 0
      nfH2O 11 46.2 46.2
      Total 20 136 136
    3. Aliquot 15 uL master mix to appropriate tubes
    4. Aliquot 4 uL reaction to appropriate tubes: use QB, QC, QE, QF, XB, XC, XE, XF
    5. Use the following thermocycler program
      1. 5 min 95C
      2. Ramp to 50C at 0.2C/s
      3. 10 min 50C
      4. 1 hr 37C; When this cycle starts add 1 uL Bglii
      5. Heat kill with 65C for 20 min