This can be achieved by taping several wells together. Be certain that the taped comb clears the bottom of the gel bed. Also prepare a mini-gel to run a 5 ul aliquot to verify complete digestion and running time to obtain desired separation. After verifying complete digestion add 10 ul of 10X glycerol dye to the digest and load on gel. Run at 80V for 4-8 hours.
Inserts are now ready for ligation.
Ligation Reaction Maniatis has several complicated formulas to determine the amounts of vector and insert to use for the most efficient ligations. The more DNA used the more efficient the ligation will be. Insert is usually the limiting factor but try to have between 100-300 ng of vector. The sample with added ligase will have bands of higher molecular weight if a substantial amount of ligation has occurred. Molecular Cloning A Laboratory Manual.