pGEM-T easy vector gives extra band

Susanne Rohrer srohrer.nospam at immv.unizh.ch
Wed Mar 7 03:04:48 EST 2001



"R. Jayakumar" wrote:

> hi..
>    I have these PCR fragment of 200bp in a pGEMT easy vector in JM109
> strain.  After digesting it with HindIII/pstI , I get an additional 1 kb
> fragment other than my linearised vector and the 200bp insert.

Doesn't that appear to be the terminally supercoiled plasmids that are not
digestible? There have been several discussions about that on this NG.

>  I have not tried fresh ligation with fresh pGEMT again, just
> because, we are running very much low on pGEMT and do not have enough money
> to buy a new stock.

uhm, how about prepping some yourself?

Susanne






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