Does CIAP cause ligation problems?

Francois Coulier coulier at infobiogen.fr
Fri Jan 10 03:00:03 EST 1997


On 9 Jan 1997, Fiberhead wrote:

> >Hi everybody,
> >
> >For the last two months I am trying to perform a non-directed ligation. 
> >Therefore I tread my vector with CIAP (Promega). However, till now I
> >wasn't 
> >successful.
> >
> 
> For what its worth, it has been my experience that inactivation of CIAP can
> be the culprit.  I think we routinely heat at 75 degrees in the presence of
> EDTA for at least 10 minutes AND phenol extract.
> 
> -seth
> 
> 
> 
> 
Boehringer Manheim is now selling shrimp alkaline phosphatase, which is
heat sensitive. This may help getting rid of it.
I have not tried it yet.
I have no affiliation with BM.

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Francois Coulier
INSERM Unite 119
27 bd Lei Roure
13009 Marseille
France

tel 33 (0) 4 91 75 84 11
fax 33 (0) 4 91 26 03 64)
email: coulier at infobiogen.fr
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