AlanFung:LabNotes/Sequencing/2009-12-5: Difference between revisions
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==Dilute libraries to 10nM== | |||
{| {{table}} | |||
| Dilute to 10nM||nM||To 10nM||ddH20||Sample | |||
|- | |||
| Dpn-CV-F2 ||26||10||8||5 | |||
|- | |||
| PGP8D||24.65||10||7.33||5 | |||
|- | |||
| PGP8E||27.17||10||8.59||5 | |||
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|} | |||
==Dilute libraries to 1nM== |
Revision as of 23:10, 5 December 2009
qPCR Quantification
PGP4 PGP6 PGP8 PGP9 PGP8D PGP8E Dpn-CV-iF2 Dpn-CV-F2
Setup | ||||
Amp Phi-X (1nM) | PGP6 | PGP9 | PGP8E | Dpn-CV-iF2 (1/10) |
Amp Phi-X (1nM) | PGP6 | PGP9 | PGP8E | Dpn-CV-iF2 (1/10) |
Amp Phi-X (1nM) | PGP6 | PGP9 | PGP8E | Dpn-CV-iF2 (1/10) |
Amp Phi-X (1nM) | PGP6 | PGP9 | PGP8E | Dpn-CV-iF2 (1/10) |
PGP4 | PGP8 | PGP8D | Dpn-CV-F2 (1/10) | |
PGP4 | PGP8 | PGP8D | Dpn-CV-F2 (1/10) | |
PGP4 | PGP8 | PGP8D | Dpn-CV-F2 (1/10) | |
PGP4 | PGP8 | PGP8D | Dpn-CV-F2 (1/10) | |
x18 | *1.1 | ||
DNA | 2 | ||
Syb_FP5 (100uM) | 0.2 | 3.6 | 3.96 |
Syb_RP7 (100uM) | 0.2 | 3.6 | 3.96 |
2X Phusion | 25 | 450 | 495 |
50X SYBR Green I | 0.2 | 3.6 | 3.96 |
H20 | 24 | 432 | 475.2 |
Total | 982.08 | ||
- 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
Average | Difference | 2^ | nM | |||||
Amp Phi-X (1nM) | 9.833 | 9.899 | 9.819 | 9.990 | 9.89 | 0 | ||
PGP4 | 9.471 | 9.522 | 9.562 | 9.706 | 9.57 | 0.32 | 1.25 | 1.25 |
PGP6 | 10.667 | 10.911 | 11.011 | 10.86 | 0.97 | 1.96 | 0.51 | |
PGP8 | 9.719 | 9.813 | 10.088 | 10.166 | 9.95 | 0.06 | 1.04 | 0.96 |
PGP9 | 9.663 | 9.710 | 9.751 | 9.71 | 0.18 | 1.13 | 1.13 | |
PGP8D | 5.250 | 5.262 | 5.287 | 5.27 | 4.62 | 24.65 | 24.65 | |
PGP8E | 5.067 | 5.143 | 5.168 | 5.13 | 4.76 | 27.17 | 27.17 | |
Dpn-CV-F2 (1/10) | 8.303 | 8.403 | 8.663 | 8.666 | 8.51 | 1.38 | 2.6 | 26 |
Dpn-CV-iF2 (1/10) | 9.824 | 9.962 | 10.000 | 9.93 | 0.04 | 1.03 | 9.74 | |
Dilute libraries to 10nM
Dilute to 10nM | nM | To 10nM | ddH20 | Sample |
Dpn-CV-F2 | 26 | 10 | 8 | 5 |
PGP8D | 24.65 | 10 | 7.33 | 5 |
PGP8E | 27.17 | 10 | 8.59 | 5 |