Alice:LabNotes/2009-9-7

From ZhangLabWiki
Jump to navigation Jump to search

Re-amplification of PhiX control library[edit]

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 EB.
PAGE gel result:
File:ZhangLab 2 2009-09-09 16hr 02minphix.jpg
QPCR result: 1.59nM