Im doing a ligation of a 6kb vector and 1kb insert. If my vector is cut with two different enzymes (they produce sticky ends) would i still need to do dephosphorylation?
Also which controls do i add for ligation. I know positive control would be a cut vector to see if ligase works. My negative control is it vector DNA and ligase but no insert? how is it different from Vector DNA no ligase and no insert? or should i do both negative controls
thanks in advance