efficient blunt ligation?

Dan Johnson djohnson at biost1.thi.tmc.edu
Fri Jun 23 19:11:13 EST 1995


In article <s-siepka-2306951435240001 at claytond2.life.uiuc.edu>,
s-siepka at uiuc.edu (Sandy Siepka) wrote:

.
> 
> Be sure to phosphatase your vector to death before attempting
> ligation--perform a control ligation and transformation first with your
> phosphatased vector alone.  This will suggest to you when your vector will
> no longer produce many 'insert-less' transfectants.
> 
Be really careful when phosphatasing your vector "to death". I found that with  
NEB  CIP overphosphatased vector does not produce any intermolecular
products. However, being modest helped. I never had problems with
blunt-end ligations since I figured an optimal amount of CIP.

M. Alexeyev



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