plasmid excision problem

Duncan Clark blackhole at
Wed Aug 10 03:22:07 EST 2005

Historians believe that in newspost 
<1123603206.895174.27540 at> on Tue, 9 Aug 
2005, vagos <el36 at> penned the following literary masterpiece:
>Do you have any ideas why this might happen and how I can avoid it?

It sounds like you are transfecting the ligation mix?

Why can't you transfect one of the correct plasmids already passaged 
through E.coli first?

If you are already doing that then methylation probably isn't the key as 
the whole plasmid will be methylated identically i.e. dam+dcm+  if in a 
standard lab E.coli.

I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.

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