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Simultaneous RNA–DNA FISH

  • Lan-Tian Lai
  • Zhenyu Meng
  • Fangwei Shao
  • Li-Feng Zhang
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1402)

Abstract

A highly useful tool for studying lncRNAs is simultaneous RNA–DNA FISH, which reveals the localization and quantitative information of RNA and DNA in cellular contexts. However, a simple combination of RNA FISH and DNA FISH often generates disappointing results because the fragile RNA signals are often damaged by the harsh conditions used in DNA FISH for denaturing the DNA. Here, we describe a robust and simple RNA–DNA FISH protocol, in which amino-labeled nucleic acid probes are used for RNA FISH. The method is suitable to detect single-RNA molecules simultaneously with DNA.

Key words

Fluorescence in situ hybridization (FISH) Single-molecule RNA FISH Amino-labeled probes Simultaneous RNA–DNA FISH lncRNA 

Notes

Acknowledgement

L-FZ is supported by the Singapore National Research Foundation under its Cooperative Basic Research Grant administered by the Singapore Ministry of Health’s National Medical Research Council. FS is supported by Nanyang Assistant Professor Fellowship (M4080531) and Ministry of Education Tier 2 Research Grant (M4020163). The authors would also like to acknowledge the Nanyang Technological University College of Science Collaborative Research Award.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.School of Biological SciencesNanyang Technological UniversitySingaporeSingapore
  2. 2.Division of Chemistry and Biological Chemistry, School of Physical and Mathematical SciencesNanyang Technological UniversitySingaporeSingapore

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