gonzales at camelot.rect.ucv.ve
Wed May 15 15:59:57 EST 2002
I have been using the erase-a-base kit from promega to delete part of a gene
I am interested in. The kit uses ExoIII to delete from a NotI site, and then
the reaction is incubated with S1 nuclease, klenow, ligase, and transformed.
I have digested my plasmid originally with NotI (Exo III sensitive) and SacI
(resistant 3´overhangs). I am now screening the colonies I have got, and I
am not sure if the SacI place has been regenerated or not. It seems likely
that the restriction site is lost throw incubation with S1 nuclease, but it
would be helpful to know it for sure. Does anybody know the answer?
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