Standard cycling mode (primer Tm ³ 60°C)Step Temperature Duration CyclesUDG activation 50°C 2 minutes HoldDual-Lock™ DNA polymerase 95°C 2 minutes HoldDenature 95°C 15 seconds 40Anneal/extend 60°C 1 minuteTable 5 Standard cycling mode (primer Tm <60°C)Step Temperature Duration CyclesUDG activation 50°C 2 minutes HoldDual-Lock™ DNA polymerase 95°C 2 minutes HoldDenature 95°C 15 seconds40Anneal 55–60°C[1] 15 secondsExtend 72°C 1 minute[1] Anneal temperature should be set to the melting point for your primers