Detecting sRNAs by Northern Blotting
Detecting sRNAs by Northern Blotting
复制标题
DOI：
10.1007/978-1-61779-083-6_3
复制
发表时间：
2011-01-01
期刊：
影响因子：
--
通讯作者：
Lopez-Gomollon, Sara
中科院分区：
文献类型：
--
作者：
Lopez-Gomollon, Sara
小分子RNA（small RNA，sRNA）在生物体转录组和蛋白质组的修饰中起着重要作用。sRNA可以根据它们的来源和方式分类为诸如microRNA、反式作用siRNA、异染色质siRNA、Piwi相互作用RNA和天然反义siRNA的类别，尽管microRNA是最保守和研究最多的类别。sRNA的表达模式可以指示其生物学功能。北方印迹法是鉴定、验证和研究sRNA表达谱的最广泛使用的方法之一，因为它是一种定量的、相对便宜的技术，对于大多数实验室来说是容易获得的。本章描述了sRNA北方印迹分析的方案，包括RNA提取、聚丙烯酰胺凝胶电泳、杂交或使用寡核苷酸探针检测sRNA。
Small RNAs (sRNAs) play fundamental roles in modifying the transcriptomes and proteomes of organisms. sRNAs can be classified according to their origin and way faction into classes such as microRNAs, transacting siRNAs, heterochromatin siRNAs, Piwi-interacting RNAs, and natural antisense siRNAs, although microRNAs are the most conserved and studied class. The expression pattern of a sRNA can be indicative of its biological function. Northern blotting is one of the most widely used methods to identify, validate, and study the expression profile of sRNAs because it is a quantitative, relatively inexpensive technique that is readily available for most laboratories. This chapter describes a protocol for sRNA Northern blot analysis, which includes RNA extraction, polyacrylamide gel electrophoresis, hybridisation, or the detection of sRNA using oligonucleotide probes.