Comparison of dot-blot and northern blot hybridizations in the determination of genetic relatedness of United States bluetongue virus serotypes

R. E. Unger, R. Y. Chuang, L. F. Chuang, R. H. Doi, Bennie Osburn

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Dot-blot and Northern blot hybridization methods to determine the genetic relatedness of United States bluetongue virus serotypes 2, 10, 11, 13, and 17 were compared. Both plasmid and insert DNA probes from cloned BTV-17 dsRNA segments 2, 5, 6, and 8 were hybridized to dsRNA from the BTV serotypes and epizootic hemorrhagic disease virus (EHDV). Stringencies of hybridization were kept identical, and experiments differed only in the method in which dsRNA was applied to the membranes (dot-blot or Northern blot). The Northern blot hybridization method yielded more consistent results, and it was visually and unequi-, vocally shown to which dsRNA segment a cDNA probe bound, since the dsRNA segments were separated by PAGE prior to blotting and hybridization. In contrast, the dot-blot hybridization method gave less consistent results. Identical results for plasmid and insert probes were obtained for Northern blot hybridizations but not for dot-blot hybridizations. At least two probes could be used simultaneously in Northern blot hybridizations.

Original languageEnglish (US)
Pages (from-to)273-282
Number of pages10
JournalJournal of Virological Methods
Volume22
Issue number2-3
DOIs
StatePublished - Jan 1 1988

Keywords

  • Dotblot hybridization
  • Genetic relatedness
  • Northern blot hybridization
  • United States bluetongue virus (BTV) serotypes

ASJC Scopus subject areas

  • Virology

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