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 Citations (Scopus)

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

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Bluetongue virus
Double-Stranded RNA
double-stranded RNA
rapid methods
Genome
genome
cell culture
gels
RNA
Gels
silver nitrate
slabs
acrylamides
Cell Culture Techniques
gel electrophoresis
Silver Nitrate
methodology
Ethidium
Acrylamide
Viral RNA

ASJC Scopus subject areas

  • Microbiology
  • veterinary(all)

Cite this

Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus. / Squire, K. R.E.; Chuang, R. Y.; Osburn, Bennie; Knudson, D. L.; Doi, R. H.

In: Veterinary Microbiology, Vol. 8, No. 6, 01.01.1983, p. 543-553.

Research output: Contribution to journalArticle

Squire, K. R.E. ; Chuang, R. Y. ; Osburn, Bennie ; Knudson, D. L. ; Doi, R. H. / Rapid methods for comparing the double-stranded RNA genome profiles of bluetongue virus. In: Veterinary Microbiology. 1983 ; Vol. 8, No. 6. pp. 543-553.
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