改良RNA提取法及樱桃PDV和PNRSV的RT-PCR检测(英文)

• 作 者: 阮小凤 Jelkmann Wilhelm 马锋旺
• 机 构: 西北农林科技大学园艺学院,Federal Biological Research Center for Agriculture and Forestry,Institute for Plant Pro
• 出 处: 西北农林科技大学学报(自然科学版)
• 发表日期: 2004年08期
• 关键词: RNA提取 RT-PCR检测 PNRSV PDV 一年生枝 芽组织 核酸提取 感病植株 有机溶剂 试剂盒

介绍了一种从木本植物组织中获得高质量RNA的快速、简单和高效的核酸提取方法。该方法是基于核酸的二氧化硅捕获,避免了使用苯酚、氯仿等有机溶剂。利用该方法从樱桃组织中提取的总RNA用RT-PCR技术检测PDV,PNRSV均获得成功。从感病植株的一年生枝的叶片、韧皮部及芽组织中扩增出了预期的目的片段,即172和449bp,而健康组织中无此扩增带。该法提取的总RNA用于RT-PCR技术检测,其敏感性至少与商业出售的QiagenRNeasy提取试剂盒相当,但简单经济。

A rapid, simple and efficient nucleic acid extraction method for obtaining high quality RNA from woody tissue is presented. The method is based on nucleic acid capture of silica and avoids the use of organic solvents such as phenol and chloroform. The total RNA extracted from cherry tissue by this method was successfully used to detect PDV and PNRSV by RT-PCR. The expected target fragments, 172 and 449 bp, were amplified from the leaves, phloem and bud tissue of one-year-old branches of susceptible plants without any amplification bands in healthy tissues. This method extracts total RNA for RT-PCR detection and is at least as sensitive as the commercially available Qiagen RNeasy kit, but simple and economical.