title: Fluorescence-based automated DNA sequencing by limited primer labeling.
abstract: The applicability of automated DNA sequencing systems to sequencing strategies that require a large number of primers is limited by the necessity for expensive fluorescence-labeled oligonucleotides. Here we present a simple procedure that allows the use of unlabeled oligonucleotides to perform fluorescence-based DNA sequencing. This method is based on a limited primer extension that incorporates three deoxynucleotides, one of which carries a fluorescent moiety. The elongated fluorescent primer is then used in a standard T7 sequencing reaction. This labeling procedure is both economical and straightforward and offers a valid alternative to current fluorescence-labeling protocols. Results of this method with different DNA templates demonstrate the reliability of the protocol.
authors.affiliation: Consiglio Nazionale delle Ricerche, Istituto di Tecnologie Biomediche, Avanzate, LITA, Segrate, Italy.
authors.forename: G
authors.initials: G
authors.lastname: De Bellis
authors.forename: R
authors.initials: R
authors.lastname: Pergolizzi
authors.forename: S
authors.initials: S
authors.lastname: Debernardi
authors.forename: L
authors.initials: L
authors.lastname: Invernizzi
authors.forename: M
authors.initials: M
authors.lastname: Luzzana
mesh_terms: D017931:DNA Primers / Q000378:metabolism*; D005453:Fluorescence; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D017931:DNA Primers