PCR A/T CLONING VECTOR

kitchingman at mbcf.stjude.org kitchingman at mbcf.stjude.org
Fri Dec 23 08:42:17 EST 1994


In article <3d9pgc$eoo at unicorn.nott.ac.uk>, Simon Dawson <mbxspd at unicorn.nott.ac.uk> writes:
> I think all you need to do is incubate Taq pol with your linearized
> plasmid for 30 mins at 70C in standard Taq buffer using 2mM dTTP
> alone.....see Marchuk et al, Nucl. Acids Res. (1991) Vol. 19(5) 
> pp 1154.
>   Hope thats of some use....by the way, if you get it working reliably
> well, can you send me any tricks you might use as I havnt tried it
> yet!
> 
> Simon Dawson
> Dept. Biochem,
> Queen's Medical Centre,
> Nottingham,
> U.K.
> 
> Internet email: mbxspd at unicorn.nott.ac.uk
> 
> ps. There IS a vector available (another NAR reference which I forget)
> which can be cut with just XcmI to give T overhangs on either end.
> If I find the reference for it, I will post it tomorrow.
The vector is pDK101 and it is available from ATCC (ATCC 77406).  The reference
is NAR 19: 4560.



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