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In situ hybridization for mRNA detection in Arabidopsis tissue sections


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.
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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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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).

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