Beware pGEM-T

Anthony Tomlinson tomlinson at pplros.demon.co.uk
Thu Sep 22 10:27:07 EST 1994


In article <pdpte-150994152610 at ppddpz.nottingham.ac.uk> Phil Emery,
pdzpte at pdn1.gene. nottingham.ac.uk writes:
In article <pdpte-150994152610 at ppddpz.nottingham.ac.uk> Phil Emery,
pdzpte at pdn1.gene. nottingham.ac.uk writes:
>If you're having trouble with Promega's pGEM-T cloning kit producing lots
>of non-recombinants, don't despair: try miniprepping a blue colony &
>cutting it with Eco RV.
>
>We did this & concluded we've been sold a tube of Eco RV-digested
>pGEM5zf(+) (as the Technical Bulletin describes), but without the 3' Ts
>added.

There do seem to be a number of untailed molecules in the pGEM-T
preparations, but there also seem to be a number of blunt PCR products in
the average Taq reaction; we haven't found pGEM-T to be too bad at PCR
cloning, especially given the blu/white screen, but I suppose you could
have a bad batch....

Anthony



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