Histo‐blotting: Hybridization in situ detection of specific RNAs on tissue sections transferred on nitrocellulose

A. V. Gudkov*, K. N. Kashkin, T. E. Zaitsevskaya, S. M. Troyanovsky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

A simple and rapid variant of in situ hybridization on tissue sections (histo‐blotting) usable for detection of specific RNA distribution among tissues is proposed. Tissue sections prepared with a cryostatic microtome are placed on nitrocellulose and these “histo‐blots” are hybridized with labelled DNA or RNA probes under conditions of Northern‐blot hybridization without any particular pretreatment. Tissue specificity of the RNA distribution may be determined by comparison of autoradiograms with the histological structure of the stained section. Histological staining and light microscopy may be carried out after hybridization of histo‐blots. Hybridization in situ may be easily combined with immunostaining under conditions of immunoblotting. Application of the proposed method is shown for alfa‐fetoprotein (AFP) and endogenous provirus (ev‐3) RNA detection in rat and chicken embryos, respectively. Histo‐blotting results correlate with the distribution of given RNAs among tissues determined by independent methods. Sensitivity, specificity and resolution of histo‐blotting have been evaluated and discussed.

Original languageEnglish (US)
Pages (from-to)1052-1056
Number of pages5
JournalInternational Journal of Cancer
Volume44
Issue number6
DOIs
StatePublished - Dec 15 1989

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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