Primer extension dideoxy chain termination nucleotide sequencing of partially purified RNA virus genomes: a technique for investigating low titre viruses with extensive genome secondary structure

C. A. Gibson, L. M. Dunster, M. Bouloy, P. D. Minor, P. G. Sanders, A. D.T. Barrett

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A modified version of the primer extension dideoxy chain termination nucleotide sequencing technique (Sanger et al., 1977) is described. This method has advantages over existing molecular cloning and primer extension techniques in that it allows the genome of RNA viruses to be directly sequenced from partially purified RNA preparations. Thus, viruses growing at unacceptably low titres in tissue culture can now be partially purified from infected mouse brain and sequenced. The technique also incorporates steps for the denaturation of secondary structure which has previously provided difficulties for primer extension sequencing.

Original languageEnglish (US)
Pages (from-to)167-176
Number of pages10
JournalJournal of Virological Methods
Volume29
Issue number2
DOIs
StatePublished - Aug 1990
Externally publishedYes

Keywords

  • Nucleotide sequencing
  • Primer extension
  • RNA virus

ASJC Scopus subject areas

  • Virology

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