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通过原位杂交和Northern杂交检测编码受体的mRNA。

Detection of mRNA encoding receptors by in situ and Northern hybridization.

作者信息

Princivalle Alessandra P, Parker Rachel M C, Dover Terri J, Barnes Nicholas M

机构信息

Biomedical Research Centre, Sheffield Hallam University, Howard Street, Sheffield, UK.

出版信息

Methods Mol Biol. 2012;897:261-302. doi: 10.1007/978-1-61779-909-9_13.

Abstract

The ability to measure mRNA encoding protein is attractive, since, provided certain criteria are met, the investigator can be confident of highly selective detection. In addition, changes in mRNA expression as well as their precise cellular localization, provides the scientist with important information that may not be evident by the detection of the translated protein. Hence, the combined approach of assessing mRNA and protein expression will often allow a more precise hypothesis to be formulated. In the present chapter, we describe methods we have optimized for the detection of mRNA following extraction from tissue or cells (Northern hybridization) and the detection of specific mRNA transcripts within their synthesising cells (in situ hybridization).

摘要

测量编码蛋白质的mRNA的能力很有吸引力,因为只要满足某些标准,研究人员就可以确信能进行高选择性检测。此外,mRNA表达的变化及其精确的细胞定位,为科学家提供了重要信息,而这些信息通过检测翻译后的蛋白质可能并不明显。因此,评估mRNA和蛋白质表达的联合方法通常能使我们更精确地提出假设。在本章中,我们描述了我们优化的方法,用于从组织或细胞中提取后检测mRNA(Northern杂交)以及在其合成细胞内检测特定的mRNA转录本(原位杂交)。

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