Uncut vector DNA-again

Nikolai Chitaev nikolaic at VISAR.WUSTL.EDU
Sat Apr 27 18:55:04 EST 1996

Shanthi Adimoolam <shanth at students.uiuc.edu> wrote:
> Hi netters,
> In my previous posting on the uncut vector problem, i forgot to state that
> i was doing this in order to avoid vector purification from the gel,
> becasue for some reason, the ligation efficiency seems really low when i
> purify the vector form the gel that i get no transformants at all. SO i
> decided to prevent gel elution by digestion with KpnI/PstI and treatment
> with CIAP to prevent its own insert (wildtype) from ligating back, but when i
> throw in the mutant insert, hopefully that will go in becasue it still has
> its phosphate groups.
> I am still unable to solve the problem of uncut vector left over.
> Please
> help!!

   It`s hard to find a gray cat in a dark room, when she isn`t there. :-) 

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