Chunxin Wang wrote:
> Then someone suggested the excess CIP might be the problem because I used
> one ul of the undiluted CIP(10u/ul) from Biolabs for dephosphorylation. I
> don't believe it because the CIP was finally inactivated by heat and
> extracted by phenol. It should have nothing to do with the packaging.
You should probably use much less than that (I think it should be diluted
10X). It's not the deactivating that the problem, too much CIP messes up you
DNA (it damages the ends of you DNA), and the damaged ends won't ligate. I
don't use CIP myself, more trouble than it is worth.
>> Any suggestion is higly appreciated.