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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

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Summary

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.

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References

  • Bonner JJ, Pardue ML (1976) Ecdysone-stimulated RNA synthesis in imaginal discs of Drosophila melanogaster. Assay by in situ hybridization. Chromosoma 58:87–99

    Google Scholar 

  • Brahic M, Haase AT (1978) Detection of viral sequences of low reiteration frequency by in situ hybridization. Proc Natl Acad Sci USA 75:6125–6129

    Google Scholar 

  • Dunn AR, Gallimore PH, Jones KW, McDougall JK (1973) In situ hybridization of Adenovirus RNA and DNA. II. Detection of adenovirus-specific DNA in transformed and tumour cells. Int J Cancer 11:628–636

    Google Scholar 

  • Godard CM, Jones KW (1979) Detection of AKR MuLV-specific RNA in AKR mouse cells by in situ hybridization. Nucleic Acids Res 6:2849–2861

    Google Scholar 

  • Godard CM, Jones KW (1980) Improved method for detection of cellular transcripts by in situ hybridization: Detection of poly(A) sequences in individual cells. Histochemistry 65:291–300

    Google Scholar 

  • Moar MH, Jones KW (1975) Detection of virus-specific DNA and RNA base-sequences in individual cells transformed or infected by adenovirus Type 2. Int J Cancer 16:998–1007

    Google Scholar 

  • Monsan P, Puzo G, Mazarguil H (1975) Etude du mécanisme d'établissement des liaisons glutaraldehyde-protéines. Biochimie 57:1281–1292

    Google Scholar 

  • Saule S, Roussel M, Lagrou C, Stéhelin D (1981) Characterization of the oncogene (erb) of avian erythroblastosis virus and its cellular progenitor. J Virol 38:409–419

    Google Scholar 

  • Schincariol AL, Joklik WK (1973) Early synthesis of virus-specific RNA and DNA in cells rapidly transformed with Rous sarcoma virus. Virology 56:532–548

    Google Scholar 

  • Taylor JM, Illmensee R, Summers J (1976) Efficient transcription of RNA into DNA by avian sarcoma virus polymerase. Biochim Biophys Acta 442:324–330

    Google Scholar 

  • Vogt VM (1973) Purification and further properties of single-strand-specific nuclease from Aspergillus Oryzae. Eur J Biochem 33:192–200

    Google Scholar 

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Authors and Affiliations

  1. Institut de Cancérologie et d'Immunogénétique (INSERM U50), Hôpital Paul Brousse, 14 et 16 Avenue Paul Vaillant Couturier, F-94800, Villejuif, France

    C. M. Godard

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  1. C. M. Godard
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Godard, C.M. Improved method for detection of cellular transcripts by in situ hybridization. Histochemistry 77, 123–131 (1983). https://doi.org/10.1007/BF00496643

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  • DOI: https://doi.org/10.1007/BF00496643

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