Daniel:Notebook/ComboLock/2017-4-12

From ZhangLabWiki
Revision as of 15:08, 12 April 2017 by >Djacobse (Created page with "=RCA Double Deuce= Back to Calendar ==Protocol== <ol> <li>Bglii digestion</li> <ol type="A"> <li>Make the following reactions; '''Do not add B...")
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
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 (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