Your lab partner says they don’t think you should use the double digested pML104 for ligation with the gRNA oligos because it won’t work as it hasn’t been cut by both enzymes. Do you agree? Why?

A) Yes, because double digestion is necessary for successful ligation.
B) No, because single digestion is sufficient for successful ligation.
C) Yes, because double digestion reduces the chances of off-target effects.
D) No, because double digestion can lead to loss of efficiency in the ligation process.