Two-color in situ hybridization: A technique for simultaneous detection of transcripts from different loci

Taiyo Toriba, Hiro Yuki Hirano

    Research output: Chapter in Book/Report/Conference proceedingChapter

    7 Citations (Scopus)

    Abstract

    Over the years, in situ hybridization has been used for visualizing spatial gene expression patterns. Direct comparison of the expression patterns of different genes is useful for studying various biological activities in many situations. If we can distinguish signals derived from different probes in the same hybridization reaction, the localization of multiple gene transcripts can be detected simultaneously. In this chapter, we describe a two-color in situ hybridization procedure, which enables us to compare the expression patterns of two genes in a single tissue section. First, we explain how to prepare RNA probes and tissue samples. Then, we explain how to perform an in situ experiment, including pre-hybridization, hybridization, post-hybridization, and detection steps. A stepwise detection procedure is used to obtain the two color signals.

    Original languageEnglish
    Title of host publicationMethods in Molecular Biology
    PublisherHumana Press Inc.
    Pages269-287
    Number of pages19
    DOIs
    Publication statusPublished - 2018 Jan 1

    Publication series

    NameMethods in Molecular Biology
    Volume1830
    ISSN (Print)1064-3745

    Keywords

    • Alkaline phosphatase
    • DIG-labeling
    • Double-staining
    • FITC-labeling
    • Fast Red TR
    • In situ hybridization
    • NBT/BCIP
    • RNA probe
    • mRNA localization

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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  • Cite this

    Toriba, T., & Hirano, H. Y. (2018). Two-color in situ hybridization: A technique for simultaneous detection of transcripts from different loci. In Methods in Molecular Biology (pp. 269-287). (Methods in Molecular Biology; Vol. 1830). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8657-6_16