transforming with RE digest

lhyatt at hkucc.hku.hk lhyatt at hkucc.hku.hk
Wed Jul 28 00:44:49 EST 1993


Hi,
	Can anyone tell me if it is OK to transform competent coli cells
with a restriction digested sample. I have been reading the recent
correspondence about checking CIPped vector, and someone suggested digesting
the ligation product with an enzyme which will only cut the vector without
insert. I want to remove a restiction site by blunt-ending with Klenow, and in
the past I have had to screen mostly native vector colonies, and this would
remove anything which still cut with the enzyme. Do I need to heat
inactivate, or phenol extract/ ethanol ppt, the ligation/ restiction product
before transformation. (I did have the required vector, but a bag of samples
has disappeared during a freezer-defrost, and I can't face screening hundreds
of minipreps which cut AGAIN!!)



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