果树病毒RNA的二氧化硅提取法

Improved RNA Virus Extraction From Fruit Trees by Silica Capture

针对果树组织含有较高的多酚和多糖物质,研究建立了适合果树病毒日常进行大量RT-PCR检测时,病毒RNA的快速、经济、高效的提取方法。该方法是基于在适宜条件下,二氧化硅对RNA的吸附和洗脱作用,避免了常规方法使用的酚、氯仿等有机溶剂。在2 h内同时可以进行数十个样品的提取,由于提取的是总RNA,1次提取,1次反转录得到的cDNA样品可以用于各种待检病毒的PCR检测。得到的总RNA和cDNA在-20℃条件下可以长期保存备用,大大提高了核酸的提取效率。

A simple, effective and efficient RNA extraction protocol has been developed for fruit tree tissues which contain high polyphenolic compounds and polysaccharides. The extract is suitable for detecting RNA viruses by RT-PCR. The method is based on Silica capture and releasing under appropriate conditions without using organic solvent such as phenol, chloroform. Decades of samples can be extracted during 2 h. Since total RNA is isolated, once transcription, the cDNA can be used for any virus detection with specific primers by PCR. The extracted RNA and transcripted cDNA can be stored at -20℃ for long time.