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LNA-Based In Situ Hybridization Detection of mRNAs in Embryos

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In Situ Hybridization Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1211))

Abstract

In situ hybridization (ISH) in embryos allows the visualization of specific RNAs as a readout of gene expression during normal development or after experimental manipulations. ISH using short DNA probes containing locked nucleic acid nucleotides (LNAs) holds the additional advantage of allowing the detection of specific RNA splice variants or of closely related family members that differ in only short regions, creating new diagnostic and detection opportunities. Here we describe methods for using short (14–24 nt) DNA probes containing LNA nucleotides to detect moderately to highly expressed RNAs in whole chick embryos during the first 5 days of embryonic development. The protocol is easily adaptable for use with embryos of other vertebrate species.

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Acknowledgement

This work was supported by NIH grant P41HD064559 to PBA.

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Authors and Affiliations

  1. Molecular Cardiovascular Research Program, Department of Cellular and Molecular Medicine, University of Arizona, Tucson, AZ, 85724, USA

    Diana K. Darnell & Parker B. Antin

Authors
  1. Diana K. Darnell
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  2. Parker B. Antin
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Corresponding author

Correspondence to Parker B. Antin .

Editor information

Editors and Affiliations

  1. Molecular Histology, Bioneer A/S, Horsholm, Denmark

    Boye Schnack Nielsen

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Darnell, D.K., Antin, P.B. (2014). LNA-Based In Situ Hybridization Detection of mRNAs in Embryos. In: Nielsen, B. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology, vol 1211. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1459-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-1459-3_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1458-6

  • Online ISBN: 978-1-4939-1459-3

  • eBook Packages: Springer Protocols

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