AlanFung:LabNotes/Sequencing/2009-11-18: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
Line 1: | Line 1: | ||
==Re-amplification of PhiX control library== | ==Re-amplification of PhiX control library== | ||
PCR: | PCR: | ||
{| {{table}} | |||
| ||||*16||*1.1 | |||
|- | |||
| PhiX Control DNA (10nM)||0.025||0.400|| | |||
|- | |||
| Syb_FP5 (100uM)||0.200||3.200||3.520 | |||
|- | |||
| Syb_RP7 (100uM)||0.200||3.200||3.520 | |||
|- | |||
| 2X Phusion||50.000||800.000||880.000 | |||
|- | |||
| 50X SYBR Green I||0.200||3.200||3.520 | |||
|- | |||
| H20||50.000||800.000||880.000 | |||
|- | |||
| | |||
|} | |||
98c 30sec -> (98C 10sec -> 65C 20sec -> 72C 20sec) X 5 cycles -> 72C 3min -> 15C | 98c 30sec -> (98C 10sec -> 65C 20sec -> 72C 20sec) X 5 cycles -> 72C 3min -> 15C |
Revision as of 01:39, 19 November 2009
Re-amplification of PhiX control library
PCR:
*16 | *1.1 | ||
PhiX Control DNA (10nM) | 0.025 | 0.400 | |
Syb_FP5 (100uM) | 0.200 | 3.200 | 3.520 |
Syb_RP7 (100uM) | 0.200 | 3.200 | 3.520 |
2X Phusion | 50.000 | 800.000 | 880.000 |
50X SYBR Green I | 0.200 | 3.200 | 3.520 |
H20 | 50.000 | 800.000 | 880.000 |
98c 30sec -> (98C 10sec -> 65C 20sec -> 72C 20sec) X 5 cycles -> 72C 3min -> 15C
Qiaquick column purification and elute in 30ul of 0.1% Tween 20-EB.
Nanodrop result:
PAGE gel result: QPCR result: