mung bean nuclease

Greg Fransen franz at winternet.com
Fri Feb 9 01:28:20 EST 1996


Anna Blom wrote:
> 
> Hello!
> 
> I seem to have big problems with Mung Bean Nuclease digestion. It
> looks as if it degrades my
> double stranded DNA - I just wanted to chew up
> the overhang left by
> BamHI. I use very little of
> enzyme - 100 times less
> then recommended by Maniatis, but it still
> does not work. My buffer
> recipe comes also from Maniatis. Anyone having experience with this
> enzyme?

I usually do the following:

1 - Make sure the enzyme is fairly "fresh"
2 - Dilute it lower than specified in protocols
3 - Incubate at room temperature

Lately I've been using NEB enzyme and their RE buffers with MgSO4 added 
(supplied by the NEB with the nuclease).  

Good Luck!



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