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
Engineering Physics Laboratory, E.I. du Pont de Nemours & Company (Inc.), Wilmington, DE 19898.
Science. 1987 Oct 16;238(4825):336-41. doi: 10.1126/science.2443975.
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.
描述了一种基于使用一组新型的四种链终止双脱氧核苷酸的DNA测序系统，每种双脱氧核苷酸都带有一种不同的经化学调节的琥珀酰荧光素染料，可通过其荧光发射加以区分。禽成髓细胞瘤病毒逆转录酶用于改进的双脱氧DNA测序方案中，以便在一个反应混合物中产生一整套荧光标记的片段。这些DNA片段在一个测序泳道中通过聚丙烯酰胺凝胶电泳进行分离，并通过与该染料组发射特性专门匹配的荧光检测系统进行鉴定。扫描系统允许同时运行多个样品，并进行基于计算机的自动碱基序列识别。本文描述了用该系统对M13噬菌体DNA进行的序列分析。
