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In situ hybridization for mRNA detection in Arabidopsis tissue sections

Nature Protocols volume 1, pages 1462–1467 (2006)Cite this article

Abstract

Plant biology is currently confronted with an overflow of expression profile data provided by high-throughput microarray transcription analyses. However, the tissue and cellular resolution of these techniques is limited. Thus, it is still necessary to examine the expression pattern of selected candidate genes at a cellular level. Here we present an in situ mRNA hybridization method that is routinely used in the analysis of gene expression patterns. The protocol is optimized for mRNA localizations in sectioned tissue of Arabidopsis seedlings including embryos, roots, hypocotyls, young primary leaves and flowers. The detailed protocol, recommended controls and troubleshooting are presented along with examples of application. The total time for the process is 10 days.

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Figure 1: Examples of results.
Figure 2

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Acknowledgements

We are very grateful to David Smyth for providing laboratory facilities and advice during the establishment of this protocol. J.H. was supported by the Ministry of Education of the Czech Republic (LC06034, MSM0021622415), P.B.B. and J.F. by the VolkswagenStiftung and the EMBO Young Investigator Program, and E.B. by the Margarete von Wrangell-Habilitationsprogramm.

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Authors and Affiliations

  1. Center for Plant Molecular Biology, Tübingen University, Tübingen, 72076, Germany

    Philip B Brewer, Jiří Friml & Eva Benková

  2. School of Biological Sciences, Monash University, Vic, 3800, Australia

    Philip B Brewer & Marcus G Heisler

  3. Department of Functional Genomics and Proteomics, Laboratory of Molecular Plant Physiology, Masaryk University, Kamenice 5, Brno, 625 00, Czech Republic

    Jan Hejátko & Jiří Friml

  4. Division of Biology, California Institute of Technology, Pasadena, 91125, California, USA

    Marcus G Heisler

Authors
  1. Philip B Brewer
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  2. Marcus G Heisler
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  3. Jan Hejátko
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  4. Jiří Friml
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  5. Eva Benková
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Correspondence to Philip B Brewer.

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Brewer, P., Heisler, M., Hejátko, J. et al. In situ hybridization for mRNA detection in Arabidopsis tissue sections. Nat Protoc 1, 1462–1467 (2006). https://doi.org/10.1038/nprot.2006.226

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