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We report on a chemical method that allows DNA sequencing by a single reaction. It is based on treatment of 5'-end-labeled DNA with N-methylformamide in the presence of manganese. This method allows the manipulation of samples to be kept to a minimum and consists of a single chemical step that requires about 30 minutes to complete base degradation, phosphodiester bond cleavage and denaturation. Examples of one-treatment, one-lane DNA sequencing of both radioactively and fluorescently 5'-end-labeled DNAs are reported.

One-Step, One-Lane Chemical DNA Sequencing by N-Methylformamide in the Presence of Metal Ions