Wang Xiaosu, Tong Yongao, Wang Shenghua
Lab of Bio-Resources and Eco-Environment, Ministry of Education, College of Life Science, Sichuan University, Chengdu 610064, China; E-Mails:
Int J Mol Sci. 2010 Sep 7;11(9):3138-48. doi: 10.3390/ijms11093138.
Northern blot analysis is a powerful research tool for discovery, validation and expression of genes, and is currently widely used to detect microRNA (miRNA) accumulation. However, the traditional Northern blot procedure, which is based on a support membrane, is overly elaborate and time-consuming, although it is unsurpassed in accuracy for determining the sizes and amounts of multiple small RNAs sharing high sequence identity. Here we present an alternative method derived from plant miRNAs, liquid Northern hybridization, using fluorescently labeled oligonucleotide probes and characterized by simple and specific miRNA determination and quantitation. The entire detection process is completed within a few hours, and multiple miRNAs can be simultaneously detected in a single experiment.
Northern印迹分析是一种用于基因发现、验证和表达的强大研究工具,目前广泛用于检测微小RNA(miRNA)的积累。然而,传统的基于支持膜的Northern印迹方法过于繁琐且耗时,尽管在确定具有高度序列同一性的多个小RNA的大小和数量方面准确性无与伦比。在此,我们提出了一种源自植物miRNA的替代方法——液相Northern杂交,该方法使用荧光标记的寡核苷酸探针,具有简单且特异性的miRNA测定和定量的特点。整个检测过程可在数小时内完成,并且在单个实验中可以同时检测多个miRNA。