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涡虫石蜡切片的RNA原位杂交

RNA In Situ Hybridization on Planarian Paraffin Sections.

作者信息

Solana Jordi

机构信息

Department of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK.

出版信息

Methods Mol Biol. 2018;1774:393-404. doi: 10.1007/978-1-4939-7802-1_13.

Abstract

RNA in situ hybridization techniques are an important tool for the study of gene expression patterns in freshwater planarians. Here I describe a RNA in situ hybridization method on histological paraffin sections of planarian tissue. This protocol allows the visualization of gene expression at cellular or subcellular resolution. Following paraffin-embedding and sectioning of planarians, the resulting sections are hybridized with hapten-labeled RNA probes. Subsequent immunological probe detection is carried out with either chromogenic or fluorescent development. This protocol can be performed alone, or in combination with other immunodetection techniques, and represents a useful alternative to whole-mount protocols more commonly used in the community.

摘要

RNA原位杂交技术是研究淡水涡虫基因表达模式的重要工具。在此,我描述一种针对涡虫组织组织学石蜡切片的RNA原位杂交方法。该方案能够在细胞或亚细胞分辨率下实现基因表达的可视化。对涡虫进行石蜡包埋和切片后,将所得切片与半抗原标记的RNA探针杂交。随后通过显色或荧光显色进行免疫探针检测。该方案可单独执行,也可与其他免疫检测技术结合使用,是该领域更常用的整装方案的一种有用替代方法。

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