Abstract
In situ hybridization is a powerful method of examining the spatial distribution of RNA molecules at the subcellular level and serves as a basic technique in the fields of cell and developmental biology. In this technique, target RNAs are fixed in cells using formaldehyde and then hybridized to complementary probes labeled with modified nucleotides that are subsequently detected by immunohistochemical methods. Here, the procedures that are commonly used for the detection of RNA in mammalian tissues and cells are described, focusing on technical tips that improve the sensitivity, productivity, and reproducibility.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Mercer TR, Dinger ME, Mattick JS (2009) Long non-coding RNAs: insights into functions. Nat Rev Genet 10:155–159
Clemson CM et al (1996) XIST RNA paints the inactive X chromosome at interphase: evidence for a novel RNA involved in nuclear/chromosome structure. J Cell Biol 132:259–275
Hutchinson JN et al (2007) A screen for nuclear transcripts identifies two linked noncoding RNAs associated with SC35 splicing domains. BMC Genomics 8:39
Sone M et al (2007) The mRNA-like noncoding RNA Gomafu constitutes a novel nuclear domain in a subset of neurons. J Cell Sci 120:2498–2506
Wilkinson DG (1998) In situ hybridization: a practical approach. Oxford University Press, Oxford
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Nakagawa, S. (2015). Analysis of the Subcellular Distribution of RNA by Fluorescence In Situ Hybridization. In: Carmichael, G. (eds) Regulatory Non-Coding RNAs. Methods in Molecular Biology, vol 1206. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1369-5_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1369-5_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1368-8
Online ISBN: 978-1-4939-1369-5
eBook Packages: Springer Protocols