Daniel:Notebook/HiResChrPaint/2014-1-10

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Revision as of 16:51, 10 January 2014 by >Djacobse (Created page with "=Alu Control (Started 1/08/2014)= Back to Calendar ==Production qPCR== Since I didn't make enou...")
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Alu Control (Started 1/08/2014)

Back to Calendar

Production qPCR

Since I didn't make enough the first time, I'm going to do another production PCR to make more Alu probe.

1. Reaction Table
  gDNA 2X Kapa SYBR qPCR MM 100 uM AP1V6-Am 100 uM AP2V6 H20 Total Volume
per rxn 1 50 0.5 0.5 48 100
PGP1F (12.1X) 12.1 605 6.05 6.05 580.8 1210
2. Aliquot 100 uL into each of 3 qPCR tubes 
3. PCR protocol:
    i. PCR 3 min 95C
    ii. 95C 10 sec
    iii. 55C 30 sec 
    iv. 72C 20 sec
    v. plate read
    vi. goto ii x30
    vii. 72C 2 min
    viii. 16C hold
4. Pool 3 samples together each
5. Column Purification, elute with 40 uL EB
6. Measure concentrations in nanodrop