Protocol for unwinding double stranded DNA with Tte UvrD Helicase (#M1202)

  1. 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
  2. Incubate at 65°C for 10 minutes

  3. 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.