Mung Bean Nuclease problem - (Aug/08/2002 )
Has anyone used mung bean nuclease and found that instead of just degrading ssDNA overhangs from linear dsDNA, it pretty much degrades it entirely?!
I'm using NEB's MBN, and although I did use more than recommended, for a twice as long as suggested, I didn't predict that it would have this effect!
Can anyone help please?
Thanks, Chris
-Chris C-
Hi Chris,
I have been working with MBN.And I had the same problem.
The reason why the enzyme suddenly digests dsDNS is that it gets unspecific when you use a high amount of it (see Maniatis).I recommend testing the proper units as I have been doing.
Chris
-sponach1-
When I had this problem, switching to higher purity water seemed to solve the problem. Possibly due to zinc in the water?
-tfitzwater-