Protocol for unwinding double stranded DNA with Tte UvrD Helicase (#M1202)
- Prepare a 20 µl reaction as follows:
Tte UvrD Helicase 20 ng (1 μL) DNA up to 1 µg Isothermal Amplification Buffer (10X) 2 μL (1X) ATP (10 mM) 2 μL (1 mM) Nuclease-Free Water to 20 μL Incubation Temperature 65°C Incubation Time 10 min
- Incubate at 65°C for 10 minutes
- Stop reaction by heating to 80°C or addition of EDTA (to 10 mM).
Note: Unwound DNA will rapidly reanneal, so consider using a trapping or digestion approach to observe the unwound DNA product of the helicase reaction.