is there anything to be aware of when injecting PCR products for rescue expts? Do they need to be phenol:ChCl3-extracted or should they be clean enough if purified thru a spin column? what sort of conc is usual when combined with pRF4?
a spin column (QiaQuick or equivalent) should be fine. As I recall, I usually did PCR products at 10-20 ng/ul for rescue.