In vitro production of large single-stranded templates for DNA sequencing.

Straus, N A; Zagursky, R J

Straus, N A; Zagursky, R J.

Afiliación

Straus NA; Department of Botany, University of Toronto, Ontario, Canada.

Biotechniques ; 10(3): 376-84, 1991 Mar.

Article en En | MEDLINE | ID: mdl-1648367

RESUMEN

A method has been developed for the preparation of large single-stranded DNA sequencing templates from primary cloning plasmids or cosmids. The method utilizes the separate action of T7 Gene 6 exonuclease and exonuclease III to generate large quantities of single-stranded template for each strand of a large-cloned fragment. Since the procedure is intended for use on primary clones, it avoids the time-consuming subcloning steps associated with most sequencing programs. The procedure also has the advantage of avoiding clone instability problems associated with subcloning in M13.

Asunto(s)

ADN de Cadena Simple/genética; Moldes Genéticos; Secuencia de Bases; Clonación Molecular; Enzimas de Restricción del ADN; ADN Recombinante; ADN de Cadena Simple/biosíntesis; Exodesoxirribonucleasas/metabolismo; Datos de Secuencia Molecular; Plásmidos

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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Moldes Genéticos / ADN de Cadena Simple Idioma: En Revista: Biotechniques Año: 1991 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Reino Unido

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