Overview
The double-stranded DNA of recombinant plasmid, phagemid, or bacteriophage M13 replicative form DNA is digested with two restriction enzymes whose sites of cleavage both lie between one end of the target DNA and the binding site for universal primer. The enzyme that cleaves nearer the target sequence must generate either a blunt end or a recessed 3' terminus; the other enzyme must generate a four-nucleotide protruding 3' terminus. By Joseph Sambrook and David W. Russell .
Details
Details of this protocol, Generation Of Sets Of Nested Deletion Mutants With Exonuclease III (Subscription Required), are located on a web site other than Biocompare Protocols.