Ligation Product is Shoter than Vector

Ken Soderstrom soderstk at ucs.orst.edu
Fri Nov 14 14:27:29 EST 1997


Here's my guess:

For some reason HindIII isn't cutting your plasmid and it remains
supercoiled.

Here's what I would do (in order):

- Make sure HindIII should cut your construct.  Did you lose the site by
subcloning?

- Make sure your HindIII is working.  Use enough for long enough and cut
something you're sure about.  

- Give up and start from scratch - unless you're obsessed with knowing
about what weird thing happened.

Good luck,


-- 
Ken Soderstrom
soderstk at ucs.orst.edu


ma <ddsyr at PUBLIC.EAST.CN.NET> wrote in article
<346AE824.5927 at public.east.cn.net>...
> Dear netters:
> At present, I clone a PCR product. When I ligate the PCR product into
pGEM7Z vector, I got unusual result.
> It should have two kind products. One is vector plus insert, one is
vector alone. But I got the third kind
> product, its size is shorter than vector after a single enzyme cut (say
HindIII, which has unique site in
> pGEM7Z). I wonder the reason. Anyone have this kind of experience and
kindly give some information.
> Ma
> 
> 



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