Cohesive Blunt end. ligation

Colin Rasmussen crasmussen at anat.med.ualberta.ca
Thu May 25 23:44:44 EST 1995


In article <3q2uth$fis at sifon.cc.mcgill.ca>, Graham Dellaire
<popa0206 at PO-Box.McGill.CA> wrote:

> When you originally cut the molecule that is when you blunt end
> 
> 1. Cut with incompatible enzyme
> 2. Klenow
> 3. phenol/chlor, ethanol precipe at -20 c

...you actually don't need to chill DNA to -20C like many people do.  BRL
showed about 10 yrs ago that the key thing is to centriguge for >10 min. 
For less than a microgram, 15-20 is even more effective.

Colin



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