Uncut vector DNA-again

Shanthi Adimoolam shanth at students.uiuc.edu
Sat Apr 27 17:38:03 EST 1996


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!!

-shanthi




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