樱桃李属坏死环斑病毒病PCR检测技术研究

Studies on PCR dete mination cherry of prunus necrotic ring-spot virus

通过用2种RNA提取方法提取甜樱桃植株样品总RNA,并对其进行电泳分析.结果表明,用方法A提取的RNA完整性比方法B较好,出现了较为典型的RNA带型.以方法A提取的甜樱桃总RNA为模板,用设计一对特异性引物进行RT-PCR反应,可见其DNA谱带与预期的一对引物应扩增的片断吻合,说明PCR检测是特异的.通过30份样品经RT-PCR扩增有2份表现出清晰、明显的特异带,占总样品数的6.7%.

Through comparing two extracting methods of complete RNA, the results of electrophoresis that method A produced the typical RNA belt and the integrality of RNA is better than method B. Using the complete RNA as template and selected primers to RT- PCR reaction, the amplified DNA belt is the same with the DNA fragment amplified by expectant primers. It indicated that the PCR reacting system is specialized. The two of 30 examined samples produced clear and distinct belt and occupied 6.7% in total samples.