Help! Direction of ligation

Namjin Chung nc1 at acpub.duke.edu
Mon Sep 30 00:02:39 EST 1996


On 23 Sep 1996, Irina A. Libova wrote:

> 
> Hi!
> I am ligating an insert into a vector, using HindiIII site on both 
> ends.  And I need a specific orientation.  But the problem is that on 
> ALL of my clones I get the reverse orientation compared to the one I 
> need, instead of getting a random 50:50 distribution of orientations.  
> Has anybode had the same problem, and what can I do to change the 
> orientation?
> 
> 

Well, try restriction and subsequent ligation of your reverse-oriented 
recombinant DNA.  This way you will better chance of getting correct 
orientation.  Hope this helps.

************************************************************************
* Namjin Chung                             *                           *
* Program in Molecular Cancer Biology      * Email: nc1 at acpub.duke.edu *
* Duke University Medical Center, Box 3345 * Voice: +1 (919) 684-2363  *
* Durham, NC 27710                         * Fax  : +1 (919) 681-8253  *
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