Enzymes in KGB?

Duke Groebe drg at prophet.pharm.pitt.edu
Thu Oct 27 08:21:41 EST 1994


In article <389k76$pvs at arcadia.informatik.uni-muenchen.de>,
salger at wap18.zi.biologie.uni-muenchen.de (Klaus Salger) wrote:
> 
> Hi netters,
> McClelland et al. described a K-glutamate buffer (KGB) in which
> a lot of different restriction enzymes, Klenow, T4 DNA polymerase,
> and T4 DNA Ligase are active. But there are some more enzymes of
> interest, phosphatases (e.g. SAP, CIP), nucleases (e.g. mung bean, S1,
> ExoIII), other polymerases (e.g. Taq, T7 DNA polymerase), in general
> all enzymes which are used to modify DNAs and perhaps other enzymes, too.
> I would appreciate if people who tried KGB with enzymes other than the
> published ones would post their experiences.
> Thanx
>   Klaus
> 
> P.S. the reference is "McClelland et al. (1988) NAR Vol 16 No 1, p.?"
> 

It has worked extremely well for me (single, double, triple digests), even
better than the buffers that come with some of the restriction enzymes. 
I've substituted potassium acetate for potassium glutamate (same
concentration) because glutamate tends to "blow out" smaller DNA bands on
agarose gels.  Acetate runs faster and thus does not interfere with smaller
DNA bands.

Duke

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