Enzymes in KGB?

Carlisle Landel landel at helios
Mon Oct 24 08:47:47 EST 1994


In article <389k76$pvs at arcadia.informatik.uni-muenchen.de>,
Klaus Salger <salger at wap18.zi.biologie.uni-muenchen.de> 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,

CIP works *very* well in KGB--in fact, you are in serious danger of
"over-CIPing", ie, getting some nuclease activity that chews up your
ends.  Add a few units of CIP after you finish digesting and incubate for
5 more minutes at 37, then heat inactivate.  

Carlisle




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