Help with ligations

Francois Coulier coulier at infobiogen.fr
Wed Jun 12 07:04:28 EST 1996

On Tue, 11 Jun 1996, Henry Wong wrote:

> Getting no ligation whatsoever with different ligases from different
> companies (BRL, Promega, Boehringer, Pharmacia), gel purified (Sigma low
> melt), single cut vector religating on itself, blunt (A-overhangs into
> T-vectors) and sticky end ligations,addition of ATP into ligation
> reactions, ligating at 12 degree and 15 as well.  What to do, any
> suggestions?  Please help.  Oh yes, cells are competent.
> Henry Wong
> MID, Universtiy of Calgary
> email: hwong at acs.ucalgary.ca
Did you adjust the pH of your ATP? Probably you did, but I spent quite 
some time a few years ago having problems with ligation, due to this 
obvious fact: ATP is acidic!
Good luck
Francois Coulier, coulier at infobiogen.fr

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