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In the post-genomic era, it is necessary to adapt methods for gene expression and functional analyses to more high-throughput levels of processing. mRNA in situ hybridization (ISH) remains a powerful tool for obtaining information regarding a gene's temporal and spatial expression pattern and can therefore be used as a starting point to define the function of a gene or a whole set of genes. We have deconstructed 'traditional' ISH techniques described for a range of organisms and developed protocols for ISH that adapt and integrate a degree of automation to standardized and shortened protocols. We have adapted this technique as a high-throughput means of gene expression analysis on wax-embedded plant tissues and also on whole-mount tissues. We have used wax-embedded wheat grains and Arabidopsis floral meristems and whole-mount Arabidopsis roots as test systems and show that it is capable of highly parallel processing.

Original publication

DOI

10.1007/978-1-59745-427-8_12

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2009

Volume

513

Pages

229 - 242

Keywords

Arabidopsis, Flowers, Gene Expression, In Situ Hybridization, Plant Roots, Plants, RNA, Messenger, RNA, Plant, Triticum