Ligation of PvuI site

Duncan Clark Duncan at
Fri Jun 17 06:02:55 EST 1994

In article: <CrHMqK.LBp at>  maryr at (Bulletin board login) writes:
> I have been trying to set up a plasmid construct using a Pvu I site.  This 
> site does not want to ligate.  I've tried BRL PvuI with no success but, have
> had a little bit of success with PVuI from New ENg. Biolabs but, no success 
> lately even with NEB enzyme.  Any help or suggestions would be greatly 
> appreciated.  Regards, Mary

If my memory serves me correctly, although the Pvu I is insensitive to dam 
methylation as such, ligation of some Pvu I sites can be. So try using plasmid
isolated from a dam- E.coli ie JM110 or GM48 (or derivs). Alternatively try
the Pvu I iso. sold by Stratagene. It is from a Bacillus sp. and may have
different characteristics. 

Duncan Clark                        | Internet:    duncan at
GeneSys Ltd.                        | Compuserve:  100015.1406 at

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