Daniel:Notebook/HiResChrPaint/2013-10-22

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Revision as of 22:38, 22 October 2013 by >Djacobse (Created page with "=BSPP Control= Back to Calendar *For first production of BSPP control, see 09/03/2013 Since I'm...")
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BSPP Control

Back to Calendar

  • For first production of BSPP control, see 09/03/2013

Since I'm out of control group, I'm going to make some more.

qPCR

1. Reaction Table
  10 nM OligoPool 2X Kapa SYBR qPCR MM 100 uM AP1V61U 100 uM AP2V6 H20 Total Volume
per rxn 0.4 50 0.4 0.4 48.8 100
BSPP 1 (12.5x) 5 625 5 5 610 1250
BSPP 2 (12.5x) 5 625 5 5 610 1250
2. Aliquot 100 uL into each of 12 qPCR tubes 
3. PCR protocol:
    i. PCR 3 min 95C
    ii. 95C 3 sec
    iii. 55C 30 sec 
    iv. 72C 20 sec
    v. plate read
    vi. goto ii x17
    vii. 72C 2 min
    viii. 16C hold
4. Pooled 3 samples together each (300 uL each, 4 pools per set, 8 total)
5. Column Purification, elute with 40 uL EB
6. Measure concentrations in nanodrop