title: Simultaneous on-line DNA sequencing on both strands with two fluorescent dyes.
abstract: We describe an automated DNA-sequencing technique which allows both the simultaneous sequencing from the two strands of double-stranded templates and the subsequent detection of the sequencing products online and in parallel. The technique is based on hardware technology also used in the ALF DNA sequencer (Pharmacia, Uppsala). A helium-neon laser was mounted into the sequencing device additionally to the standard argon laser. Two different primers, labeled with either fluorescein or Texas red, are used in a single sequencing reaction resulting in an output of two sequences. Both sequencing products are then analyzed on-line in the same lanes of a gel. This technique is especially useful for the complete sequencing of DNA fragments up to 1 kb. High accuracy sequencing of PCR products in double-stranded form can now be accomplished in only one sequencing reaction.
authors.affiliation: European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Wiemann
authors.forename: J
authors.initials: J
authors.lastname: Stegemann
authors.forename: D
authors.initials: D
authors.lastname: Grothues
authors.forename: A
authors.initials: A
authors.lastname: Bosch
authors.forename: X
authors.initials: X
authors.lastname: Estivill
authors.forename: C
authors.initials: C
authors.lastname: Schwager
authors.forename: J
authors.initials: J
authors.lastname: Zimmermann
authors.forename: H
authors.initials: H
authors.lastname: Voss
authors.forename: W
authors.initials: W
authors.lastname: Ansorge
mesh_terms: D005456:Fluorescent Dyes; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes