Vandivier Lee E, Li Fan, Gregory Brian D
Department of Biology, University of Pennsylvania, 433 S. University Ave., Philadelphia, PA, 19104, USA.
Methods Mol Biol. 2015;1284:41-70. doi: 10.1007/978-1-4939-2444-8_3.
Empirical measurement of RNA secondary structure is an invaluable tool that has provided a more complete understanding of the RNA life cycle and functionality of this extremely important molecule. In general, methods for probing structural information involve treating RNA with either a chemical or an enzyme that preferentially targets regions of the RNA in a single- or double-stranded conformation (ssRNA and dsRNA, respectively). Here, we describe an approach that utilizes a combination of ssRNA- and dsRNA-specific nuclease (ss- and dsRNase, respectively) treatments along with high-throughput sequencing technology to provide comprehensive and robust measurements of RNA secondary structure across entire plant transcriptomes.
RNA二级结构的实证测量是一种非常宝贵的工具，它能让人们更全面地了解这个极其重要分子的RNA生命周期和功能。一般来说，探测结构信息的方法包括用化学物质或酶处理RNA，这些化学物质或酶分别优先靶向单链或双链构象的RNA区域（分别为单链RNA和双链RNA）。在这里，我们描述了一种方法，该方法利用单链RNA和双链RNA特异性核酸酶（分别为单链和双链核糖核酸酶）处理的组合以及高通量测序技术，以对整个植物转录组的RNA二级结构进行全面而可靠的测量。
