摘要
[目的]探索一种简便、有效的单头蚜虫基因组DNA提取方法。[方法]以从不同地区采集的桃蚜为试材,采用改进的KAC法和优化的KAC法提取单头桃蚜的基因组DNA,用紫外分光光度计测定所提DNA的浓度（以OD260/280表示）,用0.8%琼脂糖凝胶电泳检测所提DNA的纯度,比较2种方法的提取效果;对优化KAC法所提DNA进行PCR扩增和电泳检测。[结果]优化KAC法提取的单头蚜虫基因组DNA的OD260/280为1.6-1.9,浓度为20-50 ng/g;改进KAC法提取的单头蚜虫基因组DNA的OD260/280为1.4-1.7,浓度为8-25 ng/g;来自同一地方桃蚜的基因组DNA PCR扩增产物的电泳条带基本相同。[结论]优化改进的KAC法提取的桃蚜基因组DNA浓度和纯度均较高。
[Objective] The aim was to explore a simple and effective extraction method for extracting genome DNA of single aphid.[Method] With peach aphids from different areas as the tested materials,the improved KAC method and optimized KAC method were taken to extract the genome DNA of single aphids.The concn.of obtained DNA（OD260/280） was determined by ultraviolet spectrophotometer and its purity was determined by 0.8% agarose gel electrophoresis.The extraction effects of 2 methods were compared.The DNA obtained by optimized KAC method was amplified with PCR and detected by electrophoresis.[Result] The OD260/280 of single aphids genome DNA extracted with optimized KAC method was 1.6-1.9 and its concn.was 20-50 ng/g.The OD260/280 of single aphids genome DNA extracted with improved KAC method was 1.4-1.7 and its concn.was 8-25 ng/g.The electrophoresis strip of PCR amplification products of single aphids genome DNA from the same area was similar.[Conclusion] The concn.and purity of aphids genome DNA extracted with optimized KAC method were higher.
出处
《安徽农业科学》
CAS
北大核心
2009年第10期4394-4395,4398,共3页
Journal of Anhui Agricultural Sciences
基金
甘肃省科技厅事业费(QS031-C31-15)
关键词
基因组DNA
桃蚜
提取方法
优化
Genome DNA
Peach aphid
Extraction method
Optimization
