Brewer Philip B, Heisler Marcus G, Hejátko Jan, Friml Jirí, Benková Eva
Center for Plant Molecular Biology, Tübingen University, Tübingen 72076, Germany.
Nat Protoc. 2006;1(3):1462-7. doi: 10.1038/nprot.2006.226.
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.
植物生物学目前面临着高通量微阵列转录分析提供的表达谱数据泛滥的问题。然而，这些技术的组织和细胞分辨率是有限的。因此，仍然有必要在细胞水平上检测选定候选基因的表达模式。在这里，我们介绍一种原位mRNA杂交方法，该方法常用于基因表达模式的分析。该方案针对拟南芥幼苗（包括胚胎、根、下胚轴、幼嫩的初生叶和花）切片组织中的mRNA定位进行了优化。文中给出了详细的方案、推荐的对照和故障排除方法以及应用实例。整个过程的总时间为10天。
