Daniel:Notebook/ComboLock/2017-4-12

From ZhangLabWiki
Revision as of 15:11, 12 April 2017 by >Djacobse (→‎Protocol)
Jump to navigation Jump to search

RCA Double Deuce

Back to Calendar

Protocol

  1. Bglii digestion
    1. Make the following reactions; Do not add Bglii yet
    2. Reagent Single Rxn Vol Master Mix (13.2X)
      RCA Rxn 5 0
      Bglii Cutter Primer + (100 uM) 2 26.4
      10X Buffer 3.1 2 26.4
      Bglii 1 0
      nfH2O 10 132
      Total 20 184.8
    3. Aliquot 14 uL master mix into new tube
    4. Add 5 uL appropriate sample
    5. Use the following thermocycler program
      1. 2 hr 37C; When this cycle starts add 1 uL Bglii
      2. Heat kill with 65C for 20 min