Poynter Sarah J, DeWitte-Orr Stephanie J
Department of Biology, Wilfrid Laurier University, Waterloo, ON, Canada, N2L 3G1.
Departments of Health Sciences and Biology, Wilfrid Laurier University, Waterloo, ON, Canada.
Methods Mol Biol. 2017;1656:103-118. doi: 10.1007/978-1-4939-7237-1_5.
Long double-stranded (ds) RNA molecules are produced as a byproduct of viral replication. Studying virus-derived dsRNA is important for understanding virus replication, understanding host responses to virus infections, and as a diagnostic tool for virus presence and replication. Here, we describe four different techniques for visualizing dsRNA; two antibody-dependent methods (immunoblotting and immunocytochemistry), as well as two antibody-independent methods (differential digestion and acridine orange staining). The benefits and disadvantages of each technique are also discussed.
长双链(ds)RNA分子是病毒复制的副产物。研究病毒衍生的dsRNA对于理解病毒复制、宿主对病毒感染的反应以及作为病毒存在和复制的诊断工具非常重要。在这里,我们描述了四种可视化dsRNA的不同技术;两种依赖抗体的方法(免疫印迹和免疫细胞化学),以及两种不依赖抗体的方法(差异消化和吖啶橙染色)。还讨论了每种技术的优缺点。