Daniel:Notebook/ComboLock/2017-2-1: Difference between revisions

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>Djacobse
(Created page with "=RCA Time Test= Back to Calendar ==Phosphorothiorate RCA Primer== <ol> <li>Rolling Circle Amplification</li> <ol type="A"> <li>Prepare master ...")
 
>Djacobse
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<li>Add 14 uL master mix to each tube</li>
<li>Add 14 uL master mix to each tube</li>
<li>Add sample and to new reaction tubes according to following table</li>
<li>Add sample and to new reaction tubes according to following table</li>
<li>Incubate at 30C for 12 hours</li>
 
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center"
| width="85" height="45" | Sample Code
| width="85" | Phi29 Source
| width="98" | Incubation Time (hrs)
| width="65" | Sample (1AB)
| width="65" | No C Probes (3AB)
 
|- style="font-size:12pt" align="center"
| height="15" | N
| NEB
| 3 hrs
|style="font-weight:bold" | X
|style="font-weight:bold" | X
 
|- style="background-color:#BFBFBF;font-size:12pt" align="center"
| height="15" | O
| NEB
| 3 hrs-Epicentre
|style="font-weight:bold" | X
|style="font-weight:bold" | X
 
|}
 
<li>Incubate at 30C for 3 hours</li>
<li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol>
<li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol>



Revision as of 00:47, 2 February 2017

RCA Time Test

Back to Calendar

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