Xcm I digestion of pT-NOT vector

Dr. Duncan Clark news at hgmp.mrc.ac.uk
Fri Jun 22 07:14:28 EST 2001

In article <9gv3pu$2e6r$1 at news.adamastor.ac.za>, the eminent Johan
Burger at Adamastor wrote
>Does anyone have experience in cutting the vector pT-NOT with the enzyme Xcm
>I?  This enzyme liberates a stuffer fragment to yield a T-overhang vector
>for the cloning of PCR fragments.
>We battle to get complete digestion with this enzyme on the native vector.
>It also seems that we get digestion from only one of the Xcm I sites when we
>previously cloned a fragment into another site on the vector.

There isn't by any chance a dcm methylase site at the point of cleavage?
If so try a dcm- E.coli for the plasmid prep and curse!!!

The problem with the gene pool is that there is no lifeguard.

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382

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