贝氏肩孔南极鱼基因组DNA提取方法改进的初步研究  

Preliminary study on improved genomic DNA extraction method from Trematomus bernacchii

作　　者：顾铭悦[1] 许强华[1,2,3,4] 

机构地区：[1]上海海洋大学海洋科学学院,上海201306 [2]上海海洋大学大洋渔业资源可持续开发省部共建教育部重点实验室,上海201306 [3]农业部大洋渔业资源环境科学观测实验站,上海201306 [4]国家远洋渔业工程技术研究中心,上海201306

出　　处：《上海海洋大学学报》2013年第2期168-172,共5页Journal of Shanghai Ocean University

基　　金：国家自然科学基金重大研究计划培育项目(91131006)

摘　　要：由于南极鱼的体内存在抗冻蛋白,不同于一般鱼类,常规基因组DNA的提取较为困难。为建立南极鱼基因组DNA提取的有效方法,利用贝氏肩孔南极鱼(Trematomus bernacchii)为材料,采用改进的酚-氯仿法提取基因组DNA,与传统酚-氯仿法和快速试剂盒抽提法作对比,并通过琼脂糖凝胶电泳、紫外分光光度计测定、以及PCR扩增等手段进行基因组DNA质量的检测。结果表明:利用改进的基因组DNA提取方法,从贝氏肩孔南极鱼获得的基因组DNA,电泳条带整齐明亮,A260/A280值接近1.80,适合后续的PCR扩增实验,可满足基于PCR技术的相关分子生物学实验需要。Since lots of antifreeze proteins exist in Antarctic fishes,it is quite difficult to acquire high quality genomic DNA by using normal genomic DNA extraction method.In order to establish an effective genomic DNA extraction technique for Antarctic fish,one kind of Antarctic fish,Trematomus bernacchii,was used as the experimental material,and three different kinds of genomic DNA extraction methods（the modified phenol-chloroform method,normal phenol-chloroform method and the genomic extraction kit method） were compared.The acquired genomic DNA was appraised by spectrophotometer,agarose gel electrophoresis,and PCR amplification.The results showed that we successfully obtained high quality genomic DNA from T.bernacchii by using our modified phenol-chloroform method.The gel bands were very neat and bright,the ratio of A260/A280 was around 1.80,and the acquired genomic DNA was suitable for PCR experiments.In a word,the extracted genomic DNA by using our modified phenol-chloroform method,was in good quantity and suitable for the PCR-based molecular experiment.

关 键 词：贝氏肩孔南极鱼 基因组DNA 改进酚-氯仿法 16SrRNA 

分 类 号：S917[农业科学—水产科学]