Anyone using p123T-vector?

s060965 at AIX1.UOTTAWA.CA s060965 at AIX1.UOTTAWA.CA
Wed Jun 5 13:49:23 EST 1996


I tried making my own Xcm1 T-A vector using a protocal from Biotechnology 
(sorry I don't have the reference handy).  It didn't work well.  The 
ends- produced by Xcm1 have a really low re-ligation efficiency (less 
than 50% if I remember correctly - check NEB catelog) and of course the 
PCR ligation isn;t that good anyways.

I've had much better results using a blunt -ended Bluescript, and adding 
T's using Taq at 72C for 30 minutes.

You might also want to look at the PGEM-T vector from Promega.  They sell 
it separate from the kit which includes cells etc (increasing the cost).  
Apparently this is made by using terminal transferase ... I haven;t tried 
this yet.

Cris

On Mon, 3 Jun 1996, Akira Nabetani wrote:

> I am considering trying MoBiTec's T-A vector, vector p123T. It seems to
> be shipped as circular plasmid and the users themselves will need to
> digest it with XcmI to make as "T-A" cloning vector. If it works well, I
> will reduce the cost for experiments. (I don't have to purchase expensive
> "ready-to-use" T-A vectors, because i will amplify it!) Does anyone have
> any experience with this?
> In appriciations,
> 
> Akira Nabetani, Ph.D. (nabe at ri.ncvc.go.jp)
> 
> 



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