AlanFung:LabNotes/Sequencing/2009-11-18

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Revision as of 18:31, 19 November 2009 by >Alan6017518 (→‎Re-amplification of PhiX control library)
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Re-amplification of PhiX control library[edit]

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 X4 and elute in 30ul of 0.1% Tween 20-EB.

Nanodrop result: 14.8ng/ul

PAGE gel result:
QPCR result: