AlanFung:LabNotes/Sequencing/2009-11-27: Difference between revisions

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>Alan6017518
>Alan6017518
 
Line 33: Line 33:
*aliquot 24ul to each well
*aliquot 24ul to each well
  98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
  98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
{| {{table}}
| ||||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||||||||||
|-
|
|}

Latest revision as of 07:05, 27 November 2009

  • 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