AlanFung:LabNotes/Sequencing/2009-11-19
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Quantification of New amplified Phi-X and all libraries[edit]
- Dilute all samples by 10 fold
Samples | ||||
PhiX-Stock | CVF | PGP4 | PGP10 | PGP8C |
PhiX-Stock | CVF | PGP4 | PGP10 | PGP8C |
PhiX-Stock | 1-7 PGP1 | PGP6 | NA12878 | PGP8D |
PhiX-Stock | 1-7 PGP1 | PGP6 | NA12878 | PGP8D |
PhiX | 8 PGP1 | PGP8 | PGP8A | PGP8E |
PhiX | 8 PGP1 | PGP8 | PGP8A | PGP8E |
CVI | 9 PGP1 | PGP9 | PGP8B | N12 |
CVI | 9 PGP1 | PGP9 | PGP8B | N12 |
- split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
X20 | X1.1 | ||
PhiX Control DNA (10nM) | 1.000 | ||
Syb_FP5 (100uM) | 0.200 | 4.000 | 4.400 |
Syb_RP7 (100uM) | 0.200 | 4.000 | 4.400 |
2X Phusion | 25.000 | 500.000 | 550.000 |
50X SYBR Green I | 0.200 | 4.000 | 4.400 |
H20 | 24.000 | 480.000 | 528.000 |
Results[edit]
Result | ||||
PhiX | 9.112 | Cycle Difference | 2^ | x10 (nM) |
Amp PhiX | 6.051 | 3.062 | 8.348 | 83.484 |
CVI | 6.877 | 2.236 | 4.709 | 47.093 |
CVF | 8.155 | 0.958 | 1.942 | 19.419 |
1-7 PGP1 | 8.346 | 0.766 | 1.701 | 17.011 |
8 PGP1 | 6.510 | 2.603 | 6.073 | 60.734 |
9 PGP1 | 7.244 | 1.868 | 3.650 | 36.503 |
N12 | 7.915 | 1.198 | 2.293 | 22.934 |
PGP8B | 11.776 | 2.664 | 6.339 | 1.577 |
Dilution[edit]
Dilute to 10nM | Sample (ul) | ddH2O | |
Amp PhiX | 5.000 | 36.742 | |
CVI | 5.000 | 18.546 | |
CVF | 5.000 | 4.710 | |
1-7 PGP1 | 5.000 | 3.506 | |
8 PGP1 | 5.000 | 25.367 | |
9 PGP1 | 5.000 | 13.251 | |
N12 | 5.000 | 6.467 | |
PGP8B | 5.000 | 2.885 | 1nM |