Whole-Mount and Section In Situ Hybridization in Mouse Embryos for Detecting mRNA Expression and Localization
Whole-Mount and Section In Situ Hybridization in Mouse Embryos for Detecting mRNA Expression and Localization
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小鼠胚胎中的整体和切片原位杂交检测 mRNA 表达和定位
DOI：
10.1007/978-1-4939-7714-7_12
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发表时间：
2018
影响因子：
--
通讯作者：
Koshiba-Takeuchi Kazuko
中科院分区：
文献类型：
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作者：
Moriyama Yuuta;Koshiba-Takeuchi Kazuko;Yoshitaka Nishiyama;Yoshitaka Nishiyama;Koshiba-Takeuchi Kazuko
原位杂交被定义为了解基因（例如，mRNA，ncRNA）在组织中表达。mRNA的表达可以通过杂交、免疫反应和显色步骤检测为蓝色或深紫色信号。在各种模型动物中的全基因组方法已经彻底进行，并导致了新的研究领域，旨在揭示细胞分化和发育中的新基因功能。为了阐明基因的功能，时空基因表达分析是非常重要的。在这里，我描述了整体安装和部分原位杂交的协议，并强调优化温度的相关性，和钠浓度，在杂交缓冲液和基板，以提高信号。
In situ hybridization is defined as one of the most useful and powerful methods to know where genes (e.g., mRNA, ncRNA) of interest are expressed in tissues. Expression of mRNA can be detected as blue or dark purple signals though hybridization, immunoreaction and coloring steps. Genome-wide approaches in various model animals have been conducted thoroughly, and have led to new research areas aimed at uncovering novel gene functions in cell differentiation and development. To elucidate gene function, spatiotemporal gene expression analysis is very important. Here I describe protocols of whole-mount and section in situ hybridization, and emphasize the relevance of optimizing temperature, and sodium concentration, in hybridization buffer and substrate to improve signal.