I'm trying to subclone a contruct from pBluescript to a agrobacterium vector (this one is ~11 kb) using the sites PuvII from the pBluescript and SmaI from the agrobacterium vector (both blunt ends). the final construct will be 17 kb. I tried a lot; but I never get colonies on plate.
I'm using e.coli top10 cells and using heat-shock as transformation method.
what is going wrong? any suggestion?
thanks in advance