PCR A/T CLONING VECTOR

Randy Haun rhaun at nih.gov
Wed Dec 21 14:55:40 EST 1994


In article <3d9pgc$eoo at unicorn.nott.ac.uk> Simon Dawson <mbxspd at unicorn.nott.ac.uk> writes:
>From: Simon Dawson <mbxspd at unicorn.nott.ac.uk>
>Subject: Re: PCR  A/T CLONING VECTOR
>Date: 21 Dec 1994 17:43:40 GMT
>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.

Two such vectors are described in:

Kovalic et al., Nucl. Acid Res. 19 (1991) 4560
Mead et al., Bio/technology 9 (1991) 657-663



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