Xu Zhili, Culver Gloria M
Department of Biology, University of Rochester, Rochester, New York, USA.
Methods Enzymol. 2009;468:147-65. doi: 10.1016/S0076-6879(09)68008-3.
Chemical probing is widely used as a rapid approach for assessing RNA structure, folding, and function. In this chapter, we outline procedures for handling and using chemicals commonly used to probe nucleic acids. Detailed experimental conditions and design for footprinting and modification interference are presented herein. Protocols for RNA extraction, normalization, primer extension, and data evaluation are also provided. The methods described are designed to aid in the study of large RNAs, but with slight modifications are applicable to smaller RNAs.
化学探针法作为一种评估RNA结构、折叠和功能的快速方法被广泛应用。在本章中,我们概述了处理和使用常用于探测核酸的化学物质的程序。本文还介绍了足迹法和修饰干扰的详细实验条件及设计。同时也提供了RNA提取、标准化、引物延伸和数据评估的方案。所描述的方法旨在辅助对大型RNA的研究,但稍作修改后也适用于较小的RNA。