fill-in and blunt-end ligation

[Andrew Doherty]

Marcel Hoffmann wrote:
> 
> Dear Mari,
> 
> We have good results using the Shrimp Alkaline Phosphatase from
> boehringer. This enzyme has the advantage that it is heat inactivated
> (which is not the case with the other Phosphatases).
> 
> --
> ********************************************
> Marcel Hoffmann Ph.D.
> Dept. of Pharmacochemistry
> Vrije Universiteit Amsterdam
> de Boelelaan 1083, 1081 HV Amsterdam
> tel: +31(0)20-4447572
> fax +31(0)20-4447610
> ********************************************
We tried SIP and it didn't dephosphrylate vector after 2 hr incubation.
BM couldn't give us a reason, so we went back to CIAP. We generally gel
purify fragments prior to ligation to remove CIAP, which works very
well.

Andy D
-- 
*************************************************************
Dr Andrew Doherty		email -  a.doherty at bris.ac.uk
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK
BS8 1TD
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