In situ hybridization technique for mRNA detection in whole mount Arabidopsis samples
Language English Country England, Great Britain Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
17487180
DOI
10.1038/nprot.2006.333
PII: nprot.2006.333
Knihovny.cz E-resources
- MeSH
- RNA, Antisense chemical synthesis MeSH
- Arabidopsis chemistry MeSH
- Tissue Fixation MeSH
- In Situ Hybridization methods MeSH
- RNA, Messenger analysis MeSH
- Gene Expression Regulation, Plant MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- RNA, Antisense MeSH
- RNA, Messenger MeSH
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 hybridization method that is routinely used for the analysis of plant gene expression patterns. The protocol is optimized for whole mount mRNA localizations 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 approximately 7 d, depending on the tissue used.
References provided by Crossref.org
What Has Been Seen Cannot Be Unseen-Detecting Auxin In Vivo
PHABULOSA controls the quiescent center-independent root meristem activities in Arabidopsis thaliana
