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Simultaneous RNA–DNA FISH in Mouse Preimplantation Embryos

  • Aristea Magaraki
  • Agnese Loda
  • Joost Gribnau
  • Willy M. Baarends
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1861)

Abstract

Fluorescent in situ hybridization (FISH) is a powerful cytogenetic technique that allows the visualization and quantification of RNA and DNA molecules in different cellular contexts. In general, FISH applications help to advance research, cytogenetics, and diagnostics. DNA FISH can be applied, for example, for gene mapping and for detecting genetic aberrations. RNA FISH provides information about gene expression. However, in cases where RNA and DNA molecules need to be detected in the same sample, the result is often compromised by the fact that the tissue sample is damaged due to the multitude of processing steps that are required for each application. In addition, the sequential application of RNA and DNA FISH protocols on the same sample is very time consuming. Here we describe a brief protocol that enables the combined and simultaneous detection of Xist RNA and centromeric DNA of chromosome 6 in mouse preimplantation embryos. In addition, we describe how to generate indirect-labeled probes starting from BACs. This protocol may be applied to any combination of RNA and DNA detection.

Key words

RNA and DNA FISH Combination of RNA and DNA FISH Fluorescent in situ hybridization (FISH) Mouse preimplantation embryos Xist FISH X chromosome inactivation Long noncoding RNA FISH DNA FISH Long noncoding RNAs 

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Copyright information

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

Authors and Affiliations

  • Aristea Magaraki
    • 1
  • Agnese Loda
    • 2
    • 3
  • Joost Gribnau
    • 1
  • Willy M. Baarends
    • 1
  1. 1.Department of Developmental BiologyErasmus University Medical CenterRotterdamThe Netherlands
  2. 2.Mammalian Developmental Epigenetics groupInstitut CurieParis Cedex 05France
  3. 3.Department of Developmental BiologyErasmus University Medical CenterRotterdamThe Netherlands

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