AlanFung:LabNotes/Sequencing/2009-11-23: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
Line 71: | Line 71: | ||
| | | | ||
|} | |} | ||
Latest revision as of 18:20, 23 November 2009
Quantification of New amplified Phi-X and all libraries[edit]
- Dilute concentration by 10 fold for Har_2, Har_5
- Dilute New amplified PhiX to 1nM (Mix 2ul amp. PhiX with 164.97 ddH2O)
- Keep P3, P5, P7 and CVF_N_D as is
Samples | ||||
PhiX-Stock | 8/9 P7 | |||
PhiX-Stock | 8/9 P7 | |||
PhiX(1nM) | Har_2 | |||
PhiX(1nM) | Har_2 | |||
8/9 P3 | Har_5 | |||
8/9 P3 | Har_5 | |||
8/9 P5 | CVF_N_D | |||
8/9 P5 | CVF_N_D | |||
- split every 50ul rxn into 2 wells
98C 30 sec -> 20 cycles of (98C 10sec->64C 20sec->72C 20sec)->72C 5 min
X8 | X1.1 | ||
PhiX Control DNA (10nM) | 1.000 | ||
Syb_FP5 (100uM) | 0.200 | 1.6 | 1.76 |
Syb_RP7 (100uM) | 0.200 | 1.6 | 1.76 |
2X Phusion | 25.000 | 200 | 220 |
50X SYBR Green I | 0.200 | 1.6 | 1.76 |
H20 | 24.000 | 192 | 211.2 |
nM | sample ul | Tris-CL ul | HP3 | total | |
8/9 P7 | 8.09 | 3 | 20.06 | 1.21 | 24.27 |
CVF_N_D | 2.91 | 4 | 7.06 | 0.58 | 11.64 |
Dilute to 10nM | Sample (ul) | ddH2O | ||
8/9 P3 | 4.000 | 6.928 | 2.928 | |
8/9 P5 | 4.000 | 6.628 | 2.628 | |
Har_2 | 4.000 | 5.600 | 1.600 | |
Har_5 | 4.000 | 6.876 | 2.876 | |
CVI | 5 | - | 18.546 | |
CVF | 5 | - | 4.710 |