通过显色原位杂交检测固定细胞和组织中的单个RNA
Detection of Individual RNA in Fixed Cells and Tissues by Chromogenic ISH.
作者信息
Jiang Meng, Lin Chen, Ke Rongqin
机构信息
Center for Precision Medicine, School of Biomedical Sciences and School of Medicine, Huaqiao University, Quanzhou, Fujian, 362021, China.
出版信息
Bio Protoc. 2020 Feb 5;10(3):e3510. doi: 10.21769/BioProtoc.3510.
Abstract
Visualization of RNA molecules helps to better understand the functions of expressed genes. Currently, most conventional hybridization methods for visualization of individual RNAs are based on fluorescence detection. Herein we present a chromogenic hybridization protocol for visualization of single RNA molecules in fixed cells and tissues. The protocol is based on padlock probing and rolling circle amplification to generate detectable chromogenic signal from single RNA molecules. Chromogenic signal can avoid background autofluorescence and can be preserved for a longer period than fluorescence signal.
摘要
RNA分子的可视化有助于更好地理解已表达基因的功能。目前，大多数用于单个RNA可视化的传统杂交方法都是基于荧光检测。在此，我们提出了一种用于固定细胞和组织中单个RNA分子可视化的显色杂交方案。该方案基于锁式探针和滚环扩增，从单个RNA分子产生可检测的显色信号。显色信号可以避免背景自发荧光，并且比荧光信号保存的时间更长。
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