A System for Rapid DNA Sequencing with Fluorescent Chain-Terminating Dideoxynucleotides

Abstract

A DNA sequencing system based on the use of a novel set of four chain-terminating dideoxynucleotides, each carrying a different chemically tuned succinylfluorescein dye distinguished by its fluorescent emission is described. Avian myeloblastosis virus reverse transcriptase is used in a modified dideoxy DNA sequencing protocol to produce a complete set of fluorescence-tagged fragments in one reaction mixture. These DNA fragments are resolved by polyacrylamide gel electrophoresis in one sequencing lane and are identified by a fluorescence detection system specifically matched to the emission characteristics of this dye set. A scanning system allows multiple samples to be run simultaneously and computer-based automatic base sequence identifications to be made. The sequence analysis of M13 phage DNA made with this system is described.

Keywords

DNAFluorescenceSequencing by ligationDNA sequencingDNA nanoball sequencingDNA sequencerSingle molecule real time sequencingPolymerase chain reactionBiologySequencing by hybridizationBase pairMolecular biologyComputational biologyChemistryGeneticsGenomic libraryBase sequenceGene

MeSH Terms

AutomationAvian Myeloblastosis VirusBacteriophagesBase SequenceDNADNAViralDeoxyribonucleotidesElectrophoresisPolyacrylamide GelFluoresceinsFluorescent DyesRNA-Directed DNA PolymeraseSpectrometryFluorescenceSuccinates

Affiliated Institutions

DuPont (United States) US

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Publication Info

Year: 1987
Type: article
Volume: 238
Issue: 4825
Pages: 336-341
Citations: 848
Access: Closed

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APA Style

                            
                                
                                    James M. Prober, 
                                
                                    George L. Trainor, 
                                
                                    Rudy J. Dam
                                
                                et al.
                            
                            (1987). 
                            A System for Rapid DNA Sequencing with Fluorescent Chain-Terminating Dideoxynucleotides. 
                            Science
                            , 238
                            (4825)
                            , 336-341.
                            https://doi.org/10.1126/science.2443975
                        

Identifiers

DOI: 10.1126/science.2443975
PMID: 2443975

Data Quality

Data completeness: 81%