Double digest Sma1/Sac1

Rob Kirkpatrick kirkpat at ?cc.umanitoba.ca?
Mon Jul 6 13:03:18 EST 1998


In article <6nqn62$pb3 at is.bbsrc.ac.uk>, "Andy Bettany"
<andrew.bettany at bbsrc.ac.uk> wrote:

> Dear All,
> 
> I need to double digest a plasmid with Sac1 and Sma1. Both enzymes cut in
> the same buffer (according to Boehringer). Can I add both enzymes and
> incubate at 30C? Or do I need to do the
> Sma1 first at 30C then add the Sac1 and put at 37C?
> Thanks for your help,
> 
> --

In situations like this, we'll start the digest off with Sma1 at room temp
for an hour then add the second enzyme and put it at 37C.  Sma1 still
works at 37C, but we like to give it that optimal head-start then it can
finish off it's life at 37C.  Works like a charm!!!

Rob

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