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

Rodolfo Negri | Zendy; Giovanna Costanzo | Zendy; Raffaele Saladino | Zendy; Ernesto Di Mauro | Zendy

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

Author(s) -

Rodolfo Negri,

Giovanna Costanzo,

Raffaele Saladino,

Ernesto Di Mauro

Publication year - 1996

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/96215rr04

Subject(s) - phosphodiester bond , dna , chemistry , bond cleavage , cleavage (geology) , dna sequencing , metal ions in aqueous solution , denaturation (fissile materials) , biochemistry , ion , biology , gene , organic chemistry , nuclear chemistry , rna , paleontology , fracture (geology) , catalysis

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.

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