Triple restriction digest?

Dr. Duncan Clark Duncan at
Sat Oct 14 02:45:32 EST 2000

In article <8s82hh$e5d$1 at>, the eminent  at -
Before you buy. wrote
>Is this not a good idea/waste of enzyme?  I
>have another double digest protocol, however it requires two enzymes
>that are not compatible, and would just be a little more work (and would
>give the 3.5 and 2 kb that the triple digest would do).

Either method will work fine. Consider what happens when one purifies a
single RE from a bug with many. A crude extract has all enzyme
activities (up to 5 RE's for some bugs) so you automatically see a
multi-enzyme digest. 

If you go the latter of your methods then pick the low salt buffer
enzyme first, do the digest then spike in additional salt and do the
second digest. 

The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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