DETECTION OF INDIVIDUAL ENDOGENOUS RNA TRANSCRIPTS IN SITU USING MULTIPLE SINGLY LABELED PROBES

被引：127

作者：

Raj, Arjun ^{[1
]}

Tyagi, Sanjay ^{[2
]}

机构：

[1] Univ Penn, Dept Bioengn, Philadelphia, PA 19104 USA

[2] Univ Med & Dent New Jersey, New Jersey Med Sch, Publ Hlth Res Inst, Newark, NJ 07103 USA

来源：

METHODS IN ENZYMOLOGY, VOL 472: SINGLE MOLECULE TOOLS, PT A: FLUORESCENCE BASED APPROACHES | 2010年 / 472卷

关键词：

STOCHASTIC GENE-EXPRESSION; HYBRIDIZATION; LOCALIZATION;

D O I：

10.1016/S0076-6879(10)72004-8

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Measurements of gene expression within single cells have revealed startling variability otherwise hidden in bulk measurements. Here, we present an in situ hybridization method capable of detecting individual mRNA molecules, thus permitting the accurate quantification and localization of mRNA within fixed sample. Our in situ protocol involves probing the target mRNA using a series of singly labeled oligonucleotide probes. This method is simple to implement and is applicable to a variety of biological samples. We also discuss some aspects of image processing required for analyzing the resulting data.

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收藏

页码：365 / 386

页数：22

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