Daniel:Notebook/HiResChrPaint/2014-2-21: Difference between revisions

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#Aliquot 100 uL into each of 12 qPCR tubes  
#Aliquot 100 uL into each of 6 qPCR tubes  
#PCR protocol:
#PCR protocol:
##PCR 3 min 95C
##PCR 3 min 95C
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##72C 2 min
##72C 2 min
##16C hold
##16C hold
#Pooled 3 samples together each (300 uL each, 4 pools per set, 8 total)
#Pooled 3 samples together each (300 uL each, 2 total)
#Column Purification, elute with 40 uL EB
#Column Purification, elute with 40 uL EB
#Measure concentrations in nanodrop
#Measure concentrations in nanodrop

Revision as of 18:46, 21 February 2014

Multi-Color FISH (Started 02/08/2014)

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qPCR of BSPP

Sample:

  • BSPP 1-DMR 220K Exp 1 (10nM); 2/8/11 from Noi
  1. Reaction table
  10 nM OligoPool 2X Kapa SYBR qPCR MM 100 uM AP1V61U 100 uM AP2V6 2 mM aa-dUTP H20 Total Volume
per rxn 0.4 50 0.4 0.4 2.5 46.3 100
BSPP 1 (6.1x) 2.44 305 2.44 2.44 15.25 282.43 610
  1. Aliquot 100 uL into each of 6 qPCR tubes
  2. PCR protocol:
    1. PCR 3 min 95C
    2. 95C 3 sec
    3. 55C 30 sec
    4. 72C 20 sec
    5. plate read
    6. goto ii x17
    7. 72C 2 min
    8. 16C hold
  3. Pooled 3 samples together each (300 uL each, 2 total)
  4. Column Purification, elute with 40 uL EB
  5. Measure concentrations in nanodrop

qPCR Results

qPCR Curve

[[Image:|Image:]] Nanodrop Results