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

8 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

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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