Electrophoretically uniform fluorescent dyes for automated DNA sequencing.

TitleElectrophoretically uniform fluorescent dyes for automated DNA sequencing.
Publication TypeJournal Article
Year of Publication1996
AuthorsMetzker, ML, Lu, J, Gibbs, RA
JournalScience
Volume271
Issue5254
Pagination1420-2
Date Published1996 Mar 08
ISSN0036-8075
KeywordsAutoanalysis, Base Sequence, Boron Compounds, DNA Primers, Electrophoresis, Polyacrylamide Gel, Fluorescent Dyes, Molecular Sequence Data, Sensitivity and Specificity, Sequence Analysis, DNA, Spectrometry, Fluorescence
Abstract

A class of dyes, BODIPY fluorophores, has been identified for automated DNA sequencing that has improved spectral characteristics compared with conventional fluorescein and rhodamine dyes. Single and double BODIPY dye primers were characterized in commercially available DNA sequencers and showed uniform electrophoretic mobilities and high fluorescence intensities. The improved physical properties of BODIPY dye primers were demonstrated by direct base-calling from the unprocessed fluorescent signals and improved heterozygote analyses of mixed-base populations. The high sensitivity of BODIPY dye primers requires at least 33 percent less reagent consumed per reaction than conventional dye primers, which should affect the costs of large genome-sequencing efforts.

Alternate JournalScience
PubMed ID8596915
Grant List1 P30 HG00210 / HG / NHGRI NIH HHS / United States
1 RO1 HG00823 / HG / NHGRI NIH HHS / United States
AI07483 / AI / NIAID NIH HHS / United States