Detection of single base substitutions in influenza virus RNA molecules by denaturing gradient gel electrophoresis of RNA-RNA or DNA-RNA heteroduplexes.
Single point mutations in the NS gene of influenza virus were detected by electrophoresis of double-stranded RNA heteroduplexes in denaturing gradient gels. The heteroduplex RNAs were made by hybridization of virion RNA with SP6-derived RNA probes of varying length. Mutations located at different positions along the NS gene (890 nucleotides long) were all detected in a predictable fashion. The method of heteroduplex analysis was also successfully used in detecting single point mismatches in DNA-RNA hybrids.
D001483:Base Sequence; D002874:Chromosome Mapping; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005814:Genes, Viral; D009980:Influenza A virus / Q000235:genetics*; D009154:Mutation; D009439:Neuraminidase / Q000235:genetics; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D012330:RNA, Double-Stranded / Q000032:analysis; D012367:RNA, Viral / Q000235:genetics*; D013696:Temperature
D004279:DNA, Viral; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D009439:Neuraminidase