strange transformation results

Peter Ianakiev ianakiev at genome.wi.mit.edu
Mon Apr 29 08:26:34 EST 2002


Hi William

Have you checked your vector for contaminating nucleases? If you have some
exonuclease activity in the ligation mixture, you would get lots of blunt ended
vector that can self ligate, especially when you carry the ligation for long
time.

Hope this might help

Cheers
Peter

William Sun wrote:

> I have sometimes observed a strange phenomenon while subcloning and would
> like to get some input.  When I subclone a piece of DNA into a cloning
> vector (I usually subclone using two different restr. enzymes) and transform
> into bacteria, I include a negative control of just the cut vector without
> insert.  Once in a while, I would get more colonies on my negative control
> than experimental plate.  But when I screen colonies from my experimental
> plate, most of them are positive!  Can anyone offer an explanation?
>
> William Sun
> x3930
>
> ---




More information about the Methods mailing list