Sma I religation problem

Tim Dunn t.dunn at
Tue Jul 11 20:43:09 EST 1995

In article <3tv50s$p5g at>, mikep at wrote:

> Hello everyone,
> I have been having major troubles with religating Sma I cut pGEM.  The vector
> has been cut, and the linear plasmid gel purified using Wizard clean up
> but try as I might it wont religate to itself.  The same routine performed on
> the same plasmid with Eco RI works just fine.

Hi Mike, I'm in the next building to you!
Sma1 is a notorius "nibbler", in that it will eat back ends if you are not
very careful of the reaction conditions.  Use the bare minimum of enzyme
in the correct buffer and incubate at 25 degrees C. We have troubles with
Sma from time to time....  
Hope this is of some use.

Tim Dunn
Centre for Molecular and Cellular Biology
University of Queensland
Australia 4075
e-mail t.dunn at
phone  (07) 365 4565
fax    (07) 365 4388 

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