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MicroRNA In Situ Hybridization on Whole-Mount Preimplantation Embryos

  • Karen Goossens
  • Luc Peelman
  • Ann Van Soom
Part of the Methods in Molecular Biology book series (MIMB, volume 1211)

Abstract

Whole-mount in situ hybridization (WISH) using antisense probes is widely used to visualize RNA sequences in embryos and to determine the precise site of expression in the different cells or tissues. The target sequence is hybridized with an antisense RNA probe, followed by visual or fluorescence detection to measure the site and level of expression. However, the detection of short RNA molecules is hampered by the reduced stringency of the probes for short transcripts. Here, we describe a procedure for WISH detection of short RNA molecules, like miRNAs, in mammalian preimplantation embryos using LNA-modified probes with high sensitivity and specificity.

Key words

MicroRNA Blastocyst Embryo In situ hybridization LNA 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Karen Goossens
    • 1
  • Luc Peelman
    • 1
  • Ann Van Soom
    • 2
  1. 1.Department of Nutrition, Genetics and Ethologyy - Faculty of Veterinary MedicineGhent UniversityMerelbekeBelgium
  2. 2.Department of Reproduction, Obstetrics, and Herd Health, Faculty of Veterinary MedicineGhent UniversityMerelbekeBelgium

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