Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus

K. R.E. Squire, R. Y. Chuang, Bennie Osburn, D. L. Knudson, R. H. Doi

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Various double-stranded RNA extraction procedures, gel electrophoresis systems, and methods to detect the RNA bands in the gel were investigated to find the most rapid methods to obtain the genome profiles of bluetongue virus in small volumes (1-25 ml) of infected cell culture fluids. Rapid double-stranded RNA extraction procedures coupled with staining the acrylamide gel slabs with ethidium bromide or silver nitrate resulted in well-defined genome profiles from bluetongue virus infected cell cultures in 6-48 h. Radioactive labelling of viral RNA with 32P was time consuming, cumbersome and expensive. These techniques detect less than 0.5 μg of double-stranded RNA which can be obtained from one 1-ml well of a 24-well cluster plate of bluetongue virus infected cell monolayers. The methods were therefore suitable for rapid comparisons of the electropherotypes of multiple virus isolates.

Original languageEnglish (US)
Pages (from-to)543-553
Number of pages11
JournalVeterinary Microbiology
Volume8
Issue number6
DOIs
StatePublished - Jan 1 1983

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Fingerprint Dive into the research topics of 'Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus'. Together they form a unique fingerprint.

  • Cite this