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Combined Immunofluorescence, RNA FISH, and DNA FISH in Preimplantation Mouse Embryos

  • Ikuhiro Okamoto
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)

Abstract

Transcriptional and epigenetic dynamics of the genome occur during early development in mammals. It has been difficult to study these dynamics due to the limitation of materials and the difficulty of handling. In this chapter, we describe our attempt to apply a combination of immunofluorescence (IF), and RNA and DNA fluorescent in situ hybridization (FISH) in preimplantation mouse embryos. We have concentrated on refining these techniques to study the dynamics of X chromosome inactivation in mouse embryos. The techniques and general underlying principles described here should be applicable to other mammals of interest.

Key words

Immunofluorescence RNA FISH DNA FISH Preimplantation mouse embryos X-chromosome inactivation 

Notes

Acknowledgments

We thank E. Heard for critical reading of the manuscript.

This work was supported by funds from MEXT and JST-ERATO and by JSPS KAKENHI Grant Number JP25291076 to I.O.

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Anatomy and Cell Biology, Graduate School of MedicineKyoto UniversityKyotoJapan
  2. 2.JST, ERATOKyotoJapan

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