One-step, one-lane chemical DNA sequencing by N-methylformamide in the presence of metal ions.
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.
Negri
R
R
CNR, Rome, Italy.
Costanzo
G
G
Saladino
R
R
Di Mauro
E
E
D005559:Formamides / Q000494:pharmacology*; D008345:Manganese / Q000494:pharmacology*; D017422:Sequence Analysis, DNA / Q000379:methods*; D015032:Zinc / Q000494:pharmacology
D005559:Formamides; D008345:Manganese; D015032:Zinc; C002950:methylformamide