Daniel:Notebook/HiResChrPaint/2014-8-18

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Padlock Probes (Started 8/13/2014)[edit]

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qPCR[edit]

I need more padlock probes, so I'm going to repeat qPCR with 24x samples.

Reaction Table

Component 1X Reaction 12.2X Scaled
Padlock Probes 0.2 2.44
100 uM pAP1V41U 0.4 4.88
100 uM pAP2V4 0.4 4.88
KAPA SYBR Fast Master Mix (2X) 50 610
H2O 49 597.8
Total 100 1220
  1. Prepare master mix according to table above
  2. Add 100 uL reaction to PCR tubes (5 total)
  3. qPCR Protocol
    1. 95C 30 sec
    2. 95C 10 sec
    3. 55C 20 sec
    4. 72C 30 sec
    5. goto (3.2) 17 times
    6. 72C 2 min
    7. Hold at 16C forever
  4. Ethanol Precipitation
    1. Continued tomorrow

qPCR Results[edit]

Successful run on all lanes