Ligation of PvuI site

[Duncan Clark]

In article: <CrHMqK.LBp at magnus1.com>  maryr at magnus1.com (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
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Duncan Clark                        | Internet:    duncan at genesys.demon.co.uk
GeneSys Ltd.                        | Compuserve:  100015.1406 at compuserve.com
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