1X Helicase Buffer M:
10 mM Dithiolthreitol
10 mM Magnesium Acetate
35 mM Tris-Acetate pH 7.5 @ 25°C
3 mM ATP.
Incubate at 37ºC.
Notes:
E.coli UvrD helicase may require MutL and/or SSB for optimal performance in specific applications.
: MutL stimulates UvrD helicase activity more than tenfold by loading it onto the DNA substrate (Mechanic LE, Frankel BA, Matson SW (2000) J Biol Chem 275: 38337–38346).
: MutL and SSB are required in Helicase-Dependent Amplification using E. coli UvrD (Myriam Vincent, Yan Xu, and Huimin Kong (2004) EMBO reports Vol. 5, 795 - 800).
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