Daniel:Notebook/ComboLock/2016-7-29

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Revision as of 15:25, 29 July 2016 by >Djacobse (Created page with "=Phusion Test (Started yesterday)= Back to Calendar ==Protocol== <ol start="3"> <li>Circularization</l...")
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Phusion Test (Started yesterday)

Back to Calendar

Protocol

  1. Circularization
    1. Heat inactivate enzyme by incubating for 2 minutes at 94C
    2. Chill on ice 1 min
  2. Exonuclease Digestion
    1. Prepare exonuclease I/III mix by mixing 1 uL exonuclease I (20 units/μL) and 1 uL exonuclease III (100 units/μL) per sample
    2. Reagent Stock Concentration Reagent Added uL Added per sample Master Mix (5.1X)
      ExoI 20U/uL 20 1 5.1
      ExoIII 100 U/uL 100 1 5.1
    3. Add 2 μL of exonuclease I/III mix to reaction
    4. Mix the reaction by swirling pipette around the well 5 times
    5. Incubate reaction at 37 ºC for 2 hours
    6. Heat inactivate enzyme by incubating at 94C for 5 minutes
  3. qPCR
    1. Set up reaction according to table