In situ hybridisation technique for mRNA detection in whole mount Arabidopsis samples.
| Authors | |
|---|---|
| Year of publication | 2006 |
| Type | Article in Periodical |
| Magazine / Source | Nature Protocols |
| MU Faculty or unit | |
| 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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