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

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>Alan6017518
>Alan6017518
Line 11: Line 11:
  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


  Qiaquick column purification and elute in 30ul of EB.
  Qiaquick column purification and elute in 30ul of 0.1% Tween 20-EB.
Nanodrop result:
Nanodrop result:


  PAGE gel result:
  PAGE gel result:
  QPCR result:
  QPCR result:

Revision as of 01:36, 19 November 2009

Re-amplification of PhiX control library

PCR:
                                         x4
PhiX control DNA(10nM)      0.025ul      0.1ul   
Syb_FP5(100uM)              0.2ul        0.8ul
Syb_RP7(100uM)              0.2ul        0.8ul
2x phusion master mix       50ul         200ul
50x SYBR geen               0.2ul        0.8ul
H2O                         50ul         200ul

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: