一种球蚧类蚧虫基因组DNA的提取方法  

An isolation method of genomic DNA from coccini scale insects

作　　者：李惠萍 王静慧 张龙霞 刘海峰 刘晓琳 

机构地区：[1]山西出入境检验检疫局,太原030024

出　　处：《环境昆虫学报》2014年第2期182-187,共6页Journal of Environmental Entomology

基　　金：国家质检总局科技计划项目(2005IK064)(2009IK275)

摘　　要：本研究针对一类虫体膨大,体壁硬化,革质化或木质化的球蚧类蚧虫开展了全基因组提取方法的探讨,特别是在材料的选择和前处理过程两个关键环节进行了新的尝试,并通过目前文献报道的SDS、CTAB、TES、NaCl和TNES法进行了随后的基因组提取,运用1%琼脂糖凝胶电泳、DNA OD值测定和特定基因的PCR扩增对基因组的提取效果进行了检测,结果表明,无论是冷冻保存还是75%酒精固定液-4℃保存的球蚧样本均可用于DNA提取,选择球蚧体壁作为提取材料要比整头虫体效果好,前处理采用在少量提取缓冲液中剪碎样品的方法获得的基因组机率大,纯度高、条带整齐无拖尾,同时也证实CTAB法更适合该类蚧虫基因组的提取。Coccini scale insects belong to soft scale insects. Their body would swelled up to ball or half ball before female adults started laying eggs. Then the body wall developed to harden, leathery or lignified. So, the problem was put up to extracted genomic DNA from these insects. In this study, we have explored that sample storage, materials selection and sample pretreatment techniques could give effect on genomic DNA extraction. We also tried screening tests for continuous DNA extracting by using the SDS, the CTAB, the TES, the NaC1 and the TNES methods respectively. The quality of DNA were tested and analysed by 1% agarose gel following electrophoresis, the value of DNA OD260nm/OD280nm and the special gene amplified by PCR. The result indicated samples keeped by 75% ethanol in -4℃ for 1 -3 years could use to extracted genomic DNA successfully, the body wall of Coccini scale insects was best materials using for extracting genomic DNA, and cutting materials was better than grounding to help get good quality genomic DNA. Further, our study showed that CTAB method was obviously better and more suitable for genomic DNA extraction of these scale insects.

关 键 词：球蚧 基因组DNA DNA提取 

分 类 号：Q968.1[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]