A system for rapid DNA sequencing with fluorescent chain-terminating dideoxynucleotides
| Autor: | George L. Trainor, Frank W. Hobbs, Robert J. Zagursky, Mark Anton Jensen, James Merrill Prober, Rudy Johan Dam, Anthony Joseph Cocuzza, Kirk Baumeister, Charles William Robertson |
|---|---|
| Rok vydání: | 1987 |
| Předmět: |
DNA nanoball sequencing
DNA polymerase Deoxyribonucleotides DNA sequencing chemistry.chemical_compound symbols.namesake Automation Bacteriophages Fluorescent Dyes Sanger sequencing Avian Myeloblastosis Virus Multidisciplinary biology Base Sequence RNA-Directed DNA Polymerase Succinates Ion semiconductor sequencing DNA Fluoresceins Spectrometry Fluorescence Biochemistry chemistry DNA Viral biology.protein symbols Pyrosequencing Electrophoresis Polyacrylamide Gel Single molecule real time sequencing |
| Zdroj: | Science (New York, N.Y.). 238(4825) |
| ISSN: | 0036-8075 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|