fill-in and blunt-end ligation

Oliver Sorgenfrei oliver at
Wed May 14 03:16:14 EST 1997

Martin Offterdinger wrote:
> On 12 May 1997 22:29:18 GMT, Mari.L.Shinohara at (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.


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