In situ hybridization for mRNA detection in Arabidopsis tissue sections.
Plant biology is currently confronted with an overflow of expression profile data provided by high-throughput microarray transcription analyses. However, the tissue and cellular resolution of these techniques is limited. Thus, 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 in the analysis of gene expression patterns. The protocol is optimized for mRNA localizations in sectioned tissue of Arabidopsis seedlings including embryos, roots, hypocotyls, young primary leaves and flowers. The detailed protocol, recommended controls and troubleshooting are presented along with examples of application. The total time for the process is 10 days.
Brewer
Philip B
PB
Center for Plant Molecular Biology, Tübingen University, Tübingen 72076, Germany. philip.brewer@zmbp.uni-tuebingen.de
Heisler
Marcus G
MG
Hejátko
Jan
J
Friml
Jirí
J
Benková
Eva
E
D017360:Arabidopsis / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D017403:In Situ Hybridization / Q000379:methods*; D012333:RNA, Messenger / Q000302:isolation & purification*
D012333:RNA, Messenger