A rapid method for sequencing the 5′- and 3′-termini of double-stranded RNA viral templates using RLM-RACE

被引：77

作者：

Coutts, RHA ^{[1
]}

Livieratos, IC ^{[1
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, Dept Biol Sci, London SW7 2AZ, England

来源：

JOURNAL OF PHYTOPATHOLOGY | 2003年 / 151卷 / 09期

关键词：

RNA sequencing; RLM-RACE; dsRNA templates;

D O I：

10.1046/j.1439-0434.2003.00755.x

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

A protocol is described that permits rapid, inexpensive and reliable synthesis and amplification of 5'- and 3'-terminal cDNAs using viral double-stranded (ds) RNA templates. The method of RNA ligase-mediated amplification of cDNA ends for amplification of single-stranded RNAs (Liu and Gorovsky, 1993) was adapted for dsRNAs to generate the amplicons. These amplicons can be easily cloned and sequenced to complete the sequencing of 5'- and 3'-termini of any particular virus genome. The method is of great utility and can be used on viral genomic dsRNA or viral dsRNA replicative forms when the virion RNA is unavailable.

引用

页码：525 / 527

页数：3

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