t-tailing vector of choice

m-brazil at nimr.mrc.ac.uk m-brazil at nimr.mrc.ac.uk
Mon Sep 12 07:40:07 EST 1994


Forwarded message:
> From server-daemon at dl.ac.uk Mon Sep 12 11:40:41 1994
> Resent-From: server-daemon <server-daemon at dl.ac.uk>
> Date: 12 Sep 1994 05:51:20 GMT
> Resent-Date: Mon, 12 Sep 94 11:24:49 UT
> Message-Id: <94912112449.~INN-MYCa02909.bionet-news at dl.ac.uk>
> Resent-Message-Id: <350q8o$24d at styx.uwa.edu.au>
> Precedence: list
> From: lsellner at uniwa.uwa.edu.au (Loryn Sellner)
> Reply-To: lsellner at uniwa.uwa.edu.au (Loryn Sellner)
> Original-Sender: "bionet.molbio.methds-reagnts mail newsgroup" <server-daemon at uk.ac.dl>
> To: "bionet.molbio.methds-reagnts mail newsgroup" <bionet-news at uk.ac.dl>
> Subject: T-tailing for PCR product ligation
> Comments: List problems/queries to <biosci at dl.ac.uk>
> Comments: To mail both the group and netnews send to (methods at dl.ac.uk)
> X-Article-Number: bionet.molbio.methds-reagnts Msg # 9955
> X-Listpath: bionet-news
> X-Mailer: MXT V 12.16.1
> Sender: server-daemon at dl.ac.uk
> 
> 
> Dear netters,
> I have been reading the advice for cloning of PCR products by using the 
> TA cloning vector (or any T tailed vector). Is there a way to T tail my 
> own vector so I can clone my PCR product into the site I want, (namely a 
> blunt ended restriction site)?
> Thanks.
> Loryn
> 

Dear Loryn,

You can do that very easily. Digest your vector of choice with a
blunt cutter, and if possible heat denature the enzyme. Then incubate in a
larger volume with Taq polymerase and dTTP. 

I use this method (Marchuk et al, NAR 19, 5 , 1154) and it works well.

Hope it works for you

Melanie
(m-brazil at uk.ac.mrc.nimr)



More information about the Methods mailing list