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.