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

• Verfasst von: Medzihradszky, Anna [VerfasserIn]
• Verfasst von: Schneitz, Kay [VerfasserIn]
• Verfasst von: Lohmann, Jan U. [VerfasserIn]
• Titel: Detection of mRNA expression patterns by nonradioactive in situ hybridization on histological sections of floral tissue
• Verf.angabe: Anna Medzihradszky, Kay Schneitz, and Jan U. Lohmann
• Jahr: 2014
• Jahr des Originals: 2013
• Umfang: 19 S.
• Teil: year:2014
• Teil: pages:275-93
• Teil: extent:19
• Fussnoten: Published online: 02 December 2013 ; Gesehen am 11.05.2017
• Titel Quelle: Enthalten in: Riechmann, José LuisFlower Development
• Ort Quelle: New York, NY : Humana Press, 2014
• Jahr Quelle: 2014
• Band/Heft Quelle: (2014), Seite 275-93
• ISBN Quelle: 978-1-4614-9408-9
• DOI: doi:10.1007/978-1-4614-9408-9_14
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Arabidopsis
• Sach-SW: DIG-labelling
• Sach-SW: Flower
• Sach-SW: In situ hybridization
• Sach-SW: Light microscopy
• Sach-SW: Microscopy
• K10plus-PPN: 1558529357

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.