Team:UiOslo Norway/Experiments/PCR taq
PCR with Taq DNA Polymerase (NEB):
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Reaction Setup: 2.5 µl 10X Standard Taq Reaction buffer 2.5 µl dNTPS [2 mM] 0.5 µl Forward Primer [10 µM] 0.5 µl Reverse Primer [10 µM] 1 µl Template DNA 0.125 µl Taq DNA Polymerase 17.875 µl MilliQ Water
25 µl Total Reaction
PCR Program:
Step Temperature Time Initial Denaturation 95 °C 30 seconds /5 Minutes for Colony PCR 35 Cycles 95 °C 30 seconds 45 °C – 72 °C 60 seconds 68 °C 1 minute / 1 kb Final Extension 68 °C 5 minutes Hold 4 °C forever