Daniel:Notebook/ComboLock/2017-4-12: Difference between revisions

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>Djacobse
>Djacobse
Line 45: Line 45:
<li>Aliquot 12 uL master mix into new tube</li>
<li>Aliquot 12 uL master mix into new tube</li>
<li>Add 7 uL appropriate sample</li>
<li>Add 7 uL appropriate sample</li>
<li>Use the following thermocycler program</li>
<li>Mix with vortexer, spin down, and add 1 uL Bglii</li>
<ol type="a">
<li>Incubate 2 hr 37C</li>
<li>2 hr 37C; '''When this cycle starts add 1 uL Bglii'''</li>
<li>Heat kill with 65C for 20 min</li></ol>
<li>Heat kill with 65C for 20 min</li></ol>
</ol>
</ol>

Revision as of 15:14, 12 April 2017

RCA Double Deuce

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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 7 0
      10X Buffer 3.1 2 26.4
      Bglii 1 0
      nfH2O 10 132
      Total 20 158.4
    3. Aliquot 12 uL master mix into new tube
    4. Add 7 uL appropriate sample
    5. Mix with vortexer, spin down, and add 1 uL Bglii
    6. Incubate 2 hr 37C
    7. Heat kill with 65C for 20 min