Dephosphorylation - (Mar/06/2007 )

Hi,

I am using Sal1 and Hind III to digest my vector and insert.

Is dephosphorylation necessary before I carry out ligation?

Thanks.

hi
no as sequences generated after digestion are in opposite sense.
But for more confidence, you can dephosphorylate

no need to dephosphorylate. You have 2 different ends.

Dephosphorylation is a good if you have blunt end ligations.

Better if you use the control: vector digested plus ligase without insert.
Just to be sure both SalI and HindIII are cutting your vector.