The use of exonuclease III for preparing single stranded DNA for use as a template in the chain terminator sequencing method.

Exonuclease III, which degrades DNA 3' leads to 5' and is specific for duplex DNA, can be used to prepare single stranded DNA from linear duplexes. This is shown to be suitable as a template for use with the chain terminator DNA sequencing method of Sanger et al. [1]. Strategies are discussed for preparing single stranded DNA templates by this method and, in particular, its application to the sequence analysis of DNA cloned in plasmid vectors.