一种提取荒漠拟步甲昆虫基因组DNA的新方法

A New Method for Extracting Genomic DNA from Desert Tenebrionid Insects

昆虫基因组DNA的提取是研究昆虫功能基因的关键环节。一些荒漠昆虫个体较小,不易除去的外壳会造成多糖污染,用普通动物组织DNA提取方法效果不佳。通过紫外分光光度测定、凝胶电泳检测、PCR反应及限制性酶切分析表明,CTAB-NaCl法是一种适用于拟步甲科小个体昆虫提取基因组DNA的好方法,蛋白质、多糖及RNA影响很低,完全能够满足后续DNA研究的需求。与传统方法相比简便快速,且成本低,尤其适用于拟步甲科昆虫DNA的分离。

The successful extraction of genomic DNA is of significance in researching insect functional genes. However,for certain mini-body beetles,it is difficulty to remove body＇s crusta from the samples,and the extracted genomic DNA is possibly contaminated by amylase.The results of extracting the genomic DNA from general animal tissues of the small-size insects with the common-used methods are not so ideal.In this paper,a new method, CTAB-NaCl method,for extracting genomic DNA from insect sample is described here.The qualities of the obtained DNAs were tested with several standard analyses.The results of the genomic DNAs with different concentrations, analyzed with 260 nm/280 nm ultraviolet absorption spectrophotometer,agarose gel electrophoresis and PCR detection,show that the genomic DNAs isolated from five species of Tenebrionidae are suitable for molecular biological experiments.It is proved that this method is more advantageous than the common-used methods in simplicity, cheapness and speediness.It could be widely applied in DNA extraction in researching tenebrionid insects,especially the insects inhabiting in deserts in Xinjiang,China.