fill-in and blunt-end ligation

Oliver Sorgenfrei oliver at molgen.biologie.uni-marburg.de
Wed May 14 03:16:14 EST 1997


Martin Offterdinger wrote:
> 
> On 12 May 1997 22:29:18 GMT, Mari.L.Shinohara at dartmouth.edu (Mari L.
> Shinohara) wrote:
> 
> >all different enzymes generating  5'-protruding ends.  Then I
> >dephosphrylated only the vector fragment first, which has the Amp gene
> 
> 1) It is very important to remove ALL alkaline phosphatase (normally
> calf intestine AP) before ligating, with two phenol extractions.

I have good experience with Shrimp alcaline phosphatase which can be
heat inactivated.

Oliver



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