In situ hybridisation technique for mRNA detection in whole mount Arabidopsis samples.

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Authors

HEJÁTKO Jan BLILOU I. BREWER P. B FRIML Jiří SCHERES B. BENKOVA Eva

Year of publication 2006
Type Article in Periodical
Magazine / Source Nature Protocols
MU Faculty or unit

Faculty of Science

Citation
Field Genetics and molecular biology
Keywords in situ mRNA localization; whole mount; Arabidopsis
Description High throughput microarray transcription analyses provide us with the expression profiles for large amounts of plant genes. However, their tissue and cellular resolution is limited. Thus, for detailed functional analysis, it is still necessary to examine the expression pattern of selected candidate genes at a cellular level. Here, we present an in situ mRNA hybridisation method that is routinely used for the analysis of plant gene expression patterns. The protocol is optimised for whole mount mRNA localisations in Arabidopsis seedling tissues including embryos, roots, hypocotyls and young primary leaves. It can also be used for comparable tissues in other species. Part of the protocol can also be automated and performed by a liquid handling robot. Here we present a detailed protocol, recommended controls and troubleshooting, along with examples of several applications. The total time to carry out the entire procedure is ~7 d, depending on the tissue used.
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