Marco Cristina F, Skopelitis Damianos S, Timmermans Marja C P
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA.
Center for Plant Molecular Biology, University of Tübingen, Tübingen, Germany.
Methods Mol Biol. 2019;1932:159-173. doi: 10.1007/978-1-4939-9042-9_12.
Small RNAs have vital roles in numerous aspects of plant biology. Deciphering their precise contributions requires knowledge of a small RNA's spatiotemporal pattern of accumulation. The in situ hybridization protocol described here takes advantage of locked nucleic acid (LNA) oligonucleotide probes to visualize small RNA expression at the cellular level with high sensitivity and specificity. The procedure is optimized for paraffin-embedded plant tissue sections, is applicable to a wide range of plants and tissues, and can be completed within 2-6 days.
小RNA在植物生物学的众多方面发挥着至关重要的作用。要了解它们的确切作用，需要掌握小RNA积累的时空模式。本文所述的原位杂交方案利用锁核酸（LNA）寡核苷酸探针，在细胞水平上以高灵敏度和特异性可视化小RNA的表达。该方法针对石蜡包埋的植物组织切片进行了优化，适用于多种植物和组织，并且可以在2至6天内完成。
