Efficiency in cloning and sequencing using the single-stranded bacteriophage M13

A number of approaches to sequence DNA by the chain termination method are based on cloning into M13 phage vectors and the use of a universal primer. In this paper we investigate some of the factors which influence the speed and efficiency of these approaches. A modification of the template preparation, sequencing reaction, and gel system was used to obtain more reliable and clearer ladder gels. Redundancy and deficiency of shotgun DNA sequencing were reduced by a mapping technique and the use of synthetic primers. The mapping and cloning technique was used to organize the data entry so that the computer time necessary to reconstruct the sequence out of overlaps was reduced.