Daniel:Notebook/ComboLock/2017-2-1

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Revision as of 17:17, 3 February 2017 by >Djacobse (→‎Phosphorothiorate RCA Primer)
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RCA Time Test (Started Friday Jan 27)

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Phosphorothiorate RCA Primer

  1. Rolling Circle Amplification
    1. Prepare master mix according to table below
    2. Reagent uL Added Master Mix (8.2X)
      Template 3 NA
      RCA Primer (10 uM) 2.5 20.5
      dNTP (1 mM) 5 41
      10X Buffer 2 16.4
      Phi29 1 0
      BSA (10 mg/mL) 0.4 3.28
      nfH2O 6.1 50.02
      Total 20 131.2
    3. Add 14 uL master mix to each tube
    4. Add sample and to new reaction tubes according to following table
    5. Sample Code Phi29 Source Incubation Time (hrs) Sample (1AB) No C Probes (3AB)
      N NEB 3 hrs X X
      O NEB 3 hrs-Epicentre X X
    6. Incubate at 30C for 3 hours
    7. Incubate at 65C for 10 minutes;Hold at 10C until next step

    Continued tomorrow