2种蚜虫DNA提取方法的比较与改进研究

Comparison and Improvement of Two Genomic DNA Extraction Methods from Single Aphid

蚜虫是农业生产上的重要害虫,蚜虫分子生物学研究的快速发展对于植物-蚜虫间的互作机理探讨、抗蚜基因的克隆和抗蚜农作物品种的选育越来越重要,而简便高效的单头蚜虫基因组DNA的提取一直是蚜虫分子生物学研究中的难点。为了提高单头蚜虫DNA提取的产量和质量,通过采用改进的DNA提取方法,对2种不同提取方法(蛋白酶K法和CTAB法)的蚜虫DNA产量和质量进行了比较研究。结果表明,无论从DNA的产量和质量上,CTAB法都优于蛋白酶K法,能达到测序要求,且方法简便,但是成功率低于蛋白酶K法;蛋白酶K法提取的产量较CTAB法偏低,产量和质量均达到了测序要求,且从DNA的提取成功率上高于CTAB法。2种改进后的DNA提取方法都能得到满足测序要求的DNA,2种方法各具优缺点,可根据具体实验要求进行选择。

Aphids are major agricultural pests injuring nearly all cultivated crops. The rapid development of aphid molecular biology research has become more and more important for studies on mechanism of plantaphid interaction, aphid-resistance gene isolation and efficient method for aphid DNA extraction has been commercial hybrid breeding. However, a simple and needed for a long time in aphid molecular biology research. In order to obtain high quality and quantity DNA from a single head of aphid, two different methods （proteinase K and CTAB） of aphid DNA extraction were investigated and compared in present study. The results showed that CTAB method was generally better than proteinase K method in respect of not only DNA quantity, but also DNA quality which is qualified for genome sequencing. Comparing with CTAB method, proteinase K method produced lower DNA yielding, however the DNA quantity and quality also reached the requirement of genome sequencing, and the success rate was higher than CTAB method. Thus the two different DNA extracting methods could be selected according to different experiment designing.