通过一系列的对比实验发现,RNA电泳检测效果不仅与凝胶孔径大小和稀释与否相关,而且也与电泳时间长短和RNA上样量的多少有关。建立了一套可方便快捷地鉴定动物组织总RNA质量的方法,即在孔径宽度为7mm的1.2%琼脂糖凝胶上,仅需取3~5μg总RNA,加6×RNA上样缓冲液混合并用无RNase的灭菌双蒸水稀释到一定体积后上样,电泳15-30min即可准确地鉴定RNA的质量。经实践证明,该方法实用性强,重复性好,具有良好的推广应用价值。
A series of comparative test analysis were conducted and results showed that the effect of detection of RNA by gel electrophoresis was not only associated with comb size and RNA concentration, but also associated with migration time in gel electrophoresis and the amount of RNA to be loaded. Finally, a facile, rapid and effective method for RNA quality determination was established, namely using 1.2% agarose gel with the comb of breadth 7 mm, loaded 3-5 μg RNA, mixed with 6×RNA loading buffer and diluted with RNase-free water, followed by electrophoresis for 15-30 min, and the reality of RNA quality could be determined. This method had the characteristic of reduplicate in practice, and can be used as a good reference for researcher.