An RNA in situ hybridization protocol optimized for monocot tissue

• Published in: STAR protocols Vol. 2; no. 2; p. 100398
• Main Authors: Zöllner, Nora R., Bezrutczyk, Margaret, Laureyns, Reinout, Nelissen, Hilde, Simon, Rüdiger, Frommer, Wolf B.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18-06-2021; Elsevier
• Subjects: Cell Biology; In Situ Hybridization; In Situ Hybridization - methods; Microscopy - methods; Molecular Biology; Plant Leaves - chemistry; Plant Leaves - genetics; Protocol; RNA - analysis; RNA - chemistry; RNA - genetics; Zea mays - chemistry; Zea mays - genetics; Molecular Biology; In Situ Hybridization; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100398

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).
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•Optimized for monocot tissues and tested on maize, sorghum, and teosinte•Clear results with low background and high resolution•Three unique samples for each gene of interest increase specificity•Comprehensive protocol with detailed explanation of all necessary steps
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.