Electrophoretically uniform fluorescent dyes for automated DNA sequencing.
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.
Metzker
M L
ML
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA.
Lu
J
J
Gibbs
R A
RA
D001322:Autoanalysis; D001483:Base Sequence; D001896:Boron Compounds / Q000737:chemistry; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000737:chemistry; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013050:Spectrometry, Fluorescence
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D017931:DNA Primers; D005456:Fluorescent Dyes