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