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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-