Zhang Zuo, Carmichael Gordon G
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA.
Methods Mol Biol. 2004;257:75-84. doi: 10.1385/1-59259-750-5:075.
In this work we describe methods for the analysis of RNAs that have been edited by the double-strand RNA-specific adenosine deaminase, ADAR. These RNAs contain inosine residues that can be detected and quantified by a variety of approaches, including base hydrolysis and thin-layer chromatography, reverse transcription polymerase chain reaction, primer extension, and inosine-specific base cleavage. The most common method for the analysis of editing will be described here. This method involves complete hydrolysis of edited RNAs to nucleoside monophosphates, followed by separation of the products using thin-layer chromatography.
在本研究中,我们描述了分析经双链RNA特异性腺苷脱氨酶ADAR编辑的RNA的方法。这些RNA含有肌苷残基,可通过多种方法进行检测和定量,包括碱水解和薄层色谱法、逆转录聚合酶链反应、引物延伸以及肌苷特异性碱基切割。本文将描述分析编辑的最常用方法。该方法包括将编辑后的RNA完全水解为单磷酸核苷,然后使用薄层色谱法分离产物。