烟粉虱全基因组DNA四种提取方法的比较

Comparison of Four Methods for Whole Genomic DNA Extraction from Bemisia tabaci

获得大量、高质量的全基因组DNA是在DNA水平上研究许多生物的分子机制的基本前提。微小昆虫由于其体型微小,单头提取DNA浓度低,无法满足部分试验所需。为寻找一种方便、快捷、高效的全基因组提取方法,参考国内外单头微小昆虫基因组DNA提取常用方法,以烟粉虱为研究材料,选取醋酸钾(KAc)法、盐析法、氯仿-异戊醇法和苯酚提取法四种方法进行多头烟粉虱的全基因组DNA提取。通过微量核酸蛋白分析仪、基因组DNA直接琼脂糖凝胶电泳、甲基化敏感扩增多态性(Methylation sensitive amplification polymorphism,MSAP)引物扩增产物的琼脂糖凝胶电泳对DNA样品进行检测,比较提取DNA的产量、质量,并综合分析各提取方法所需时间及所用试剂的毒性大小。结果表明,盐析法提取的DNA平均浓度为521 ng/μL,纯度能满足分子检测要求,用MSAP引物能得到较好的扩增结果,相比其它方法,盐析法更简便、快速且无毒。因此盐析法是多头烟粉虱全基因组DNA提取的最佳方法。

Obtaining high-quality DNA is a prerequisite to study the molecular mechanisms of many organisms. For tiny insects, low DNA concentration（from one single insect）is unable to meet the need of some experiments. To find a convenient and efficient method for whiteflies genome extraction, KAc, salting-out, chloroform-isoamylol and phenol methods were used to extract bull whiteflies genome DNA, respectively. The efficiency of different DNA extraction methods were evaluated by comparing the concentration, purity, PCR products of DNA, extraction time and the toxicity of the reagents, respectively. The results showed that the average concentration of DNA extraction of salting-out method was 521 ng/μL, and the purity could meet the experiment requirements, and the PCR product brand of MSAP primer amplification was bright. Compared to other methods, salting-out method was more simple, rapid and non-toxic. Therefore, salting-out method was the best ideally method to extract the whole genomic DNA from whiteflies.