Kun:LabNotes/ASE/2007-10-19
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Exp.1 Repeat the PGP1Ver2 PCR on GM20431 gDNA, Hues6 gDNA, Hues6 cDNA[edit]
Enzyme: Qiagen HotStarPlus Taq
Reactions:
One 33x Template 1ul 33ul 10X buffer 5ul 165ul 25mM MgCl2 3ul 99ul 10mM dNTP 1ul 33ul 50X SYBG I 0.4ul 13.2ul 10uM Primers 1ul each ThermoStart Taq 0.5ul 8.25ul H2O 38.35ul 1266ul
95C 5min -> 40 cycles of (94C 30S -> 57C 30S -> 72C 1min) -> 72C 3min on Chromo4.
Still the results are not as good as the plate with Taq 2x master mix. Less than 50% of the reactions came up.