Not able to digest plasmid with NotI

Hadi hadi at cutmeout.helix.nih.gov
Wed Nov 5 23:28:44 EST 1997


In article <345E2DD8.7611 at lacdr.leidenuniv.nl>, 
santbrin at CHEM.LEIDENUNIV.NL says...
> Hello fellow netters,
> 
> Recently I ligated a synthetic linker in the XhoI/SpeI site of a
> plasmid.
> The linker contains (from 5' to 3') a XhoI-, a BamHI-, a EcoRV-,a NotI-
> and a SpeI-site. I checked the presence of the linker by digestion of
> the plasmid (isolated by a standard alkaline lysis procedure) with the
> restriction enzymes mentioned above. They all linearize the plasmid except
> Not I: 

....truncated....

> Peter van Santbrink
> University of Leiden
> Leiden, The Netherlands
> E-mail: santbrin at lacdr.leidenuniv.com
> 
>  
Hmmm....sounds weird. Personally, I had never these kind of problems with 
NotI or ligation of NotI containing linkers. But before you start 
fiddeling around, I would sequence the plasmid to verify your NotI site. 

Hadi Al-Hasani
hadi at nojunkmailsplease.helix.nih.gov



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