Detection of mRNA expression patterns by nonradioactive in situ hybridization on histological sections of floral tissue.

PMID:24395263

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Cited by: 13

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Abstract

Analysis of gene activity with high spatial resolution is a prerequisite for deciphering regulatory networks which underlie developmental programs. Over many years, in situ hybridization has become the gold standard for the identification of in vivo expression patterns of endogenous mRNAs. Nonetheless, the method has several limitations, and the detection of lowly expressed transcripts is still a challenge. Here, we present a robust protocol for sensitive analysis of expression patterns in inflorescence tissue of Arabidopsis thaliana. We describe how the samples are fixed, embedded, and sectioned in preparation for in situ hybridization, how RNA probes are prepared, and how hybridization and detection is carried out. While the described protocol is optimized for inflorescence meristems, it can possibly be used for other tissues as well.

Keywords

Gene Expression

MeSH terms

Arabidopsis

Flowers

Gene Expression Profiling

Histological Techniques

In Situ Hybridization

RNA Probes

RNA, Messenger

Tissue Embedding

Tissue Fixation

Vacuum

Authors

Medzihradszky, Anna

Schneitz, Kay

Lohmann, Jan U

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