Daniel:Notebook/HiResChrPaint/2013-10-22: Difference between revisions

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>Djacobse
(Created page with "=BSPP Control= Back to Calendar *For first production of BSPP control, see 09/03/2013 Since I'm...")
 
>Djacobse
Line 16: Line 16:
| width="65" | 10 nM OligoPool
| width="65" | 10 nM OligoPool
| width="65" | 2X Kapa SYBR qPCR MM
| width="65" | 2X Kapa SYBR qPCR MM
| width="65" | 100 uM AP1V61U
| width="65" | 100 uM AP1V61U-Amino
| width="65" | 100 uM AP2V6
| width="65" | 100 uM AP2V6
| width="65" | H20
| width="65" | H20

Revision as of 22:41, 22 October 2013

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-Amino 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