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One-Step, One-Lane Chemical DNA Sequencing by N-Methylformamide in the Presence of Metal Ions
oleh: Rodolfo Negri, Giovanna Costanzo, Raffaele Saladino, Ernesto Di Mauro
Format: | Article |
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Diterbitkan: | Future Science Ltd 1996-11-01 |
Deskripsi
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.