AlanFung:LabNotes/Sequencing/2009-11-27

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  • Ethanol Precipitation
  • Dissolve all in 14ul ddH2O

QPCR Quantification[edit]

Setup
Stock Phi-X (1nM)   CVF-N (Shearing) (1/10)
Stock Phi-X (1nM)   CVF-N (Shearing) (1/10)
Stock Phi-X (1nM)   CVF-N (Shearing) (1/10)
Stock Phi-X (1nM)   CVF-N (Shearing) (1/10)
Amp Phi-X (1nM)     CVF-N (A/B) (1/10)
Amp Phi-X (1nM)     CVF-N (A/B) (1/10)
Amp Phi-X (1nM)     CVF-N (A/B) (1/10)
Amp Phi-X (1nM)     CVF-N (A/B) (1/10)
X8 X1.1
DNA 1.000
Syb_FP5 (100uM) 0.200 1.600 1.760
Syb_RP7 (100uM) 0.200 1.600 1.760
2X Phusion 25.000 200.000 220.000
50X SYBR Green I 0.200 1.600 1.760
dH2O 24.000 192.000 211.200
  • Dilute sample, 1ul in 9ul ddh2o
  • mix 2ul 1/10 sample with 98ul mix
  • aliquot 24ul to each well
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
Sum Ave Diff 2^ X10
CVF-N-S 8.162 24.091 8.030 2.819 7.057 70.567
0.070 8.040
7.889
PhiX Stock 11.020 32.548 10.849
0.070 10.883
10.645
CVF-N A/B 7.850 32.161 8.040 1.791 3.460 34.599
0.050 8.002
8.146
8.163
PhiX Stock 9.665 29.493 9.831
0.050 9.876
9.952