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

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>Alan6017518
>Alan6017518
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*mix 2ul 1/10 sample with 98ul mix
*mix 2ul 1/10 sample with 98ul mix
*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

Revision as of 06:38, 27 November 2009

  • Ethanol Precipitation
  • Dissolve all in 14ul ddH2O

QPCR Quantification

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