Rapid Construction of Nested Deletions of Recombinant Plasmid DNA for Dideoxy Sequening

摘要

Construction of a nested set of unidirectional deletions in an efficient method for sequencing large DNA fragments (1-5). The method relies on certain nucleases, such as exonuclease III (Exo III), that digest DNA in a predictable fashion (2). The uniform digestion rate of the exonuclease generates a progressive set of deletions of increasing size from a clone containing the entire DNA fragment. One strand of the entire DNA fragment usually can be sequenced with a single sequencing primer using this procedure.