Publication Date:
1987-10-16
Description:
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.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Prober, J M -- Trainor, G L -- Dam, R J -- Hobbs, F W -- Robertson, C W -- Zagursky, R J -- Cocuzza, A J -- Jensen, M A -- Baumeister, K -- New York, N.Y. -- Science. 1987 Oct 16;238(4825):336-41.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Engineering Physics Laboratory, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/2443975" target="_blank"〉PubMed〈/a〉
Keywords:
Automation
;
Avian Myeloblastosis Virus/enzymology
;
Bacteriophages/genetics
;
*Base Sequence
;
DNA/*genetics
;
DNA, Viral/genetics
;
*Deoxyribonucleotides/adverse effects/therapeutic use
;
Electrophoresis, Polyacrylamide Gel
;
Fluoresceins
;
*Fluorescent Dyes
;
RNA-Directed DNA Polymerase/metabolism
;
Spectrometry, Fluorescence
;
Succinates
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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