AlanFung:LabNotes/Sequencing/2009-11-18: Difference between revisions

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>Alan6017518
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==Re-amplification of PhiX control library==
==Re-amplification of PhiX control library==
  PCR:
  PCR:
                                          x4
{| {{table}}
PhiX control DNA(10nM)     0.025ul      0.1ul 
| ||||*16||*1.1
Syb_FP5(100uM)             0.2ul        0.8ul
|-
Syb_RP7(100uM)             0.2ul        0.8ul
| PhiX Control DNA (10nM)||0.025||0.400||
2x phusion master mix      50ul        200ul
|-
50x SYBR geen              0.2ul        0.8ul
| Syb_FP5 (100uM)||0.200||3.200||3.520
H2O                        50ul        200ul
|-
| 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: