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Whole-Mount and Section In Situ Hybridization in Mouse Embryos for Detecting mRNA Expression and Localization

  • Kazuko Koshiba-Takeuchi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1752)

Abstract

In situ hybridization is defined as one of the most useful and powerful methods to know where genes (e.g., mRNA, ncRNA) of interest are expressed in tissues. Expression of mRNA can be detected as blue or dark purple signals though hybridization, immunoreaction and coloring steps. Genome-wide approaches in various model animals have been conducted thoroughly, and have led to new research areas aimed at uncovering novel gene functions in cell differentiation and development. To elucidate gene function, spatiotemporal gene expression analysis is very important. Here I describe protocols of whole-mount and section in situ hybridization, and emphasize the relevance of optimizing temperature, and sodium concentration, in hybridization buffer and substrate to improve signal.

Key words

In situ hybridization Whole-mount Section Mouse embryo mRNA Expression analysis 

Notes

Acknowledgments

I thank F. Nakayama and D. Matsumura for technical assistance, and J. K. Takehchi for critical reading of the manuscript and useful advice.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Faculty of Life SciencesToyo UniversityOra-gunJapan

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