5 min ligation

David Micklem drm21 at mole.bio.cam.ac.uk
Mon Jan 9 14:36:29 EST 1995



> >Dave Boyd wrote:
> >Anybody try Boehringer Mannheim's 5 minute ligation kit yet? 
> >Whats the secret ingredient? Does it really work?
> >
> >Dave Boyd
> >University of Manitoba                  

What's the big deal with this?  I've always done sticky-end ligations for
15 minutes 
at  room temperature (using NEB ligase) and they work really well.  Lots
of colonies.
Even for 3 or 4-way ligations.  I usually use 1hr for blunt end ligations
though.

I'm not sure  I'd make a library this way though - after all, they are so
much work 
that you might as well take the extra time just in case you get more
colonies.  Maybe
one day I should do a side-by-side 15min vs overnight, but since its works
so well
for the stuff I do I've never bothered.

Try it, you'll like it!

David

_____________________________________________________________
D.R.Micklem,
Wellcome/CRC Institute,       Time flies like an arrow...
Tennis Court Road,              
Cambridge CB2 1QR             Fruit flies like a banana.
UK                             
Tel: [+44] (0)223 334129      Email:drm21 at mole.bio.cam.ac.uk
Fax: [+44] (0)223 334089               
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