An RNA in situ hybridization protocol optimized for monocot tissue

Nora R Zöllner; Margaret Bezrutczyk; Reinout Laureyns; Hilde Nelissen; Rüdiger Simon; Wolf B Frommer

doi:10.1016/j.xpro.2021.100398

An RNA in situ hybridization protocol optimized for monocot tissue

STAR Protoc. 2021 Mar 17;2(2):100398. doi: 10.1016/j.xpro.2021.100398. eCollection 2021 Jun 18.

Authors

Nora R Zöllner¹, Margaret Bezrutczyk¹, Reinout Laureyns^{2

3}, Hilde Nelissen^{2

3}, Rüdiger Simon⁴, Wolf B Frommer^{1

5}

Affiliations

¹ Institute for Molecular Physiology, Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.
² Ghent University, Department of Plant Biotechnology and Bioinformatics, Ghent 9052, Belgium.
³ VIB Center for Plant Systems Biology, Ghent 9052, Belgium.
⁴ Institute for Developmental Genetics, Heinrich-Heine-University Düsseldorf, Düsseldorf 40225, Germany.
⁵ Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Nagoya 464-8601, Japan.

Abstract

RNA in situ hybridization can be time-consuming and difficult to troubleshoot. Here, we provide an optimized protocol for maize leaf tissue, though it can be applied to other plant tissues such as shoot apical meristems, embryos, and floral organs. We generate three >100 bp unique antisense probes for each gene of interest and hybridize them to tissue sections. For complete details on the use and execution of this protocol, please refer to Bezrutczyk et al. (2021).

Keywords: Cell Biology; In Situ Hybridization; Molecular Biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

In Situ Hybridization / methods*
Microscopy / methods
Plant Leaves / chemistry
Plant Leaves / genetics
RNA* / analysis
RNA* / chemistry
RNA* / genetics
Zea mays / chemistry
Zea mays / genetics*

Substances

RNA