Chemical DNA sequencing in the opposite direction to the dideoxy chain termination method, using a single preparation of M13 single-stranded DNA

Abstract

A strategy has been developed allowing the use of a single preparation of single-stranded DNA clones for chemical DNA sequencing in the opposite direction to the classical dideoxy chain termination method. Oligonucleotide complementary to the 5′-end of the multipurpose cloning sequence, with the proper restriction enzyme, is used to cleave specifically the molecules to expose a unique 5′-end, upstream to the inserted DNA, for the kinase labeling reaction. No further treatments are necessary before Maxam–Gilbert chemical sequencing reactions.