单条线虫DNA提取方法研究

Research on DNA Extraction Methods from a Single Nematode

为探寻一种高效、稳定、廉价的单条线虫DNA提取方法,以6种植物寄生线虫作为试验材料,比较了4种方法,即直接裂解法、冻融裂解法、切割裂解法、切割冻融裂解法对单条线虫DNA的提取效果。结果表明,对线虫进行管内切割的切割冻融裂解法与切割裂解法高效、稳定,冻融裂解法效果较差,直接裂解法效果最差。对线虫进行管内切割的切割冻融裂解法和切割裂解法可高效、稳定地获得高质量的单条线虫总DNA,满足下一步的PCR扩增需求。

To explore an efficient,stable,low-cost DNA extraction method from single nematode for the support of molecular identification of nematodes,using six species of plant-parasitic nematodes as experimental materials, four DNA extraction methods,namely,direct lysis method,freeze-thaw lysis method,cutting lysis method,freeze-thaw cutting lysis method were designed and compared in order to check their efficiency of DNA extraction from single nematode. The results showed that cutting lysis method and freeze-thaw cutting lysis method were the most efficient and stable methods for DNA extraction of single nematode,then the freeze-thaw lysis method,and the direct lysis method was the worst one. Total DNA with high-quality from a single nematode could be gained efficiently and stably through two methods,namely,cutting lysis method and freeze-thaw cutting lysis method. And the extracted DNA meeted requirement of the following PCR amplification.