Long Tianyun, Lu Rui
Department of Biological Sciences, Louisiana State University, 202 Life Science Building, Baton Rouge, LA, 70803, USA.
Methods Mol Biol. 2017;1656:79-88. doi: 10.1007/978-1-4939-7237-1_3.
Northern blot analysis has been widely used as a tool for detection and characterization of specific RNA molecules. When coupled with radioactive probe northern blot allows for robust detection and characterization of small RNA molecules of trace amount. Here, we describe the detection and size characterization of virus-derived small interfering RNAs (vsiRNAs) in C. elegans using nonradioactive DNA oligo probes in northern blotting. Our protocol allows for the detection and characterization of not only primary vsiRNAs but also secondary vsiRNAs, a class of single-stranded vsiRNAs that has distinct migration pattern, and can be easily adapted to the detection of vsiRNAs in other organisms.
Northern印迹分析已被广泛用作检测和鉴定特定RNA分子的工具。当与放射性探针结合使用时,Northern印迹能够可靠地检测和鉴定痕量的小RNA分子。在此,我们描述了使用Northern印迹中的非放射性DNA寡核苷酸探针检测秀丽隐杆线虫中病毒衍生的小干扰RNA(vsiRNA)并对其进行大小鉴定。我们的方案不仅可以检测和鉴定初级vsiRNA,还可以检测和鉴定次级vsiRNA,这是一类具有独特迁移模式的单链vsiRNA,并且可以很容易地适用于检测其他生物体中的vsiRNA。