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A system for rapid DNA sequencing with fluorescent chain-terminating dideoxynucleotides.
J M Prober1, G L Trainor, R J Dam
1Engineering Physics Laboratory, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
Science (New York, N.Y.)
|October 16, 1987
Summary
A novel DNA sequencing system uses four unique chain-terminating dideoxynucleotides with distinct fluorescent dyes. This method enables simultaneous sequencing and automatic base identification for efficient DNA analysis.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- DNA sequencing is crucial for genetic research and diagnostics.
- Existing methods often require multiple reactions or complex detection systems.
Purpose of the Study:
- To describe a novel, efficient DNA sequencing system.
- To introduce a new set of fluorescently labeled chain-terminators.
Main Methods:
- Utilized four chemically tuned succinylfluorescein dyes on dideoxynucleotides.
- Employed avian myeloblastosis virus reverse transcriptase in a modified dideoxy protocol.
- Resolved fragments via polyacrylamide gel electrophoresis in a single lane.
Main Results:
- Generated a complete set of fluorescence-tagged DNA fragments in one reaction.
- Achieved identification using a fluorescence detection system matched to dye emissions.
- Demonstrated simultaneous sample analysis and automatic base sequencing.
Conclusions:
- The described DNA sequencing system offers a streamlined approach.
- This method facilitates efficient and accurate DNA sequence analysis.
- The system is suitable for applications like M13 phage DNA sequencing.