Can CIP dephosphorylate 3' protruding end? - (Jun/13/2005 )

Hi! Does somebody know if calf intestine phosphatase can also dephosphorylate 3' protruding ends ? If yes, are the reaction conditions the same as for 5' dephosphorylation?

Thanks!

hi
assuming 3' termini have hydroxyle residues, there's no need to be dephosphorylate. I would ask NEB but as written in NEB catalog, CIP can dephosphorylate 5' phosphate groups...

hi fred!

actually I have 5' recessed phosphates that I want to get rid of... I cut with KpnI, which gives 3' protruding ends....
by the way: do you dilute your ligation before transformation? I read it should help...

Thanks a lot!

well i better understand your question. I think cip can do the job you need. And after dephosphorylation, i purify on gel my dna (caus i do double digest and then dephosphorylate).

I do not dilute my dna but i 'm gonna try...