AlanFung:LabNotes/Sequencing/2009-11-23: Difference between revisions
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==Quantification of New amplified Phi-X and all libraries== | ==Quantification of New amplified Phi-X and all libraries== | ||
*Dilute concentration by 10 fold for Har_2, Har_5 | *Dilute concentration by 10 fold for Har_2, Har_5 | ||
*Dilute New amplified PhiX to 1nM | *Dilute New amplified PhiX to 1nM (Mix 2ul amp. PhiX with 164.97 ddH2O) | ||
*Keep P3, P5, P7 and CVF_N_D as is | *Keep P3, P5, P7 and CVF_N_D as is | ||
{| {{table}} | {| {{table}} |
Revision as of 16:36, 23 November 2009
Quantification of New amplified Phi-X and all libraries
- 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 |