pGEM-T easy vector gives extra band

R. Jayakumar jakku at mrna.tn.nic.in
Fri Mar 9 09:40:41 EST 2001


Dear Dr. Clark
   They use dTTP along with taq polymerase at a conc. of 2mM. for 2 hours at
70C.  please refer to article in  NAR, 1990. vol19, no. 16.
   sincerely
jayakumar


----- Original Message -----
From: "Dr. Duncan Clark" <d_clark at nospam.demon.co.uk>
To: <methods at hgmp.mrc.ac.uk>
Sent: Friday, March 09, 2001 2:49 PM
Subject: Re: pGEM-T easy vector gives extra band


> In article <3AA793A5.74B141AB at bbm1.ucm.es>, the eminent Sergio at UCM
> wrote
> >Try using the terminal transferase from any vendor, adding dTTP to a
blunt end.
> >I don't have the protocol right here, but i think there is a brief one in
> >Sambrook, and in some catalogs.
>
> Won't just using TTP generate a long multi-T tail. Should one use ddTTP?
> I need to check the original references but I think it was dTTP and Taq
> or ddTTP and TdT.
>
> Duncan
> --
> The problem with being on the cutting edge is that you occasionally get
> sliced from time to time....
>
> Duncan Clark
> GeneSys Ltd.
> Tel: +44(0)1252376288
> FAX: +44(0)8701640382
> http://www.dnamp.com
> http://www.genesys.demon.co.uk
>
>
>


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