Improved method for detection of cellular transcripts by in situ hybridization. Detection of virus-specific RNA in rous sarcoma virus-infected cells by in situ hybridization to cDNA.
Optimal conditions for the detection of complex RNA sequences in individual cells by in situ hybridization have been determined by using in vitro cultured quail embryonic cells infected with Rous sarcoma virus and a single-stranded 3H-cDNA probe of high specific activity complementary to the RSV genome. It is shown that fixation of target tissue can be suitably achieved by using glutaraldehyde at low concentration, and subjecting cytological preparations to heat post-fixation treatment. Conditions for removing unhybridized radioactive probe molecules by means of S1 nuclease are reported.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D002478:Cells, Cultured; D004277:DNA, Single-Stranded; D005814:Genes, Viral; D006358:Hot Temperature; D009693:Nucleic Acid Hybridization; D011784:Quail; D012334:RNA, Neoplasm / Q000032:analysis*; D001357:Sarcoma, Avian / Q000032:analysis* / Q000382:microbiology
D004277:DNA, Single-Stranded; D012334:RNA, Neoplasm