transformation problem

Wed Jun 12 10:35:36 EST 2002

Are you sure that band is a oc form?.
A possible explanation would be that one of the enzymes is not cutting properly 
on the vector or on the insert, and what you get is the ligation product of two 
vectors, two inserts or vector+insert (linearized). As you are not getting any 
religated vector I would say that the vector restriction sites are OK. Check the 
sites on the insert.
If everything seems to be OK, I would start to think on toxicity problems of the 
cloned protein...

satya lakshmi wrote:

> hi
> iam facing problem with ligation of a cloned vector and
> insert both are PshA1/Bstap1 digested. but when i transfrom my ligated
> product i am not getting any colonies while the ligated product shows
> a band above the vector running like a relaxed circular form when
> compared with the undigested.
> can anyone help me as aim unable to understand what could be the
> possible problem?
> satya

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