摘要
鳍条是研究鱼类分子生物学常用的组织样本,长期保存的鳍条为实验提供源源不断的基因组DNA。本研究介绍一种便捷保存鱼类鳍条用于DNA提取的方法---干燥法,并说明了用此方法保存鲤(Cyprinus carpio)、施氏鲟(Acipenser schrenckii)和虹鳟(Onchorynchus mykiss)鳍条效果。采集新鲜鳍条,贴在滤纸等吸水性强的纸张上,覆盖另一张滤纸,防止受潮、霉变,自然干燥后在室温保存3个月。剪取部分鳍条样本,用常规酚、氯仿抽提法提取基因组DNA,分别用紫外分光光度计、琼脂糖凝胶电泳、PCR扩增方法检测基因组DNA的质量。结果显示:用干燥法保存的3种鱼类的鳍条样品,提取获得的基因组DNA的OD260/OD280值在1.82-1.89之间, OD260/OD230值在2.29-2.70之间,琼脂糖凝胶电泳条带清晰明亮,可以用于后续的分子生物学研究。本研究证实用干燥法长期保存鱼类鳍条能够获得高质量的基因组DNA,为珍贵样品的长期保存提供了一种行之有效的方法,也为远距离、大样品的采集和保存提供了便捷的选择。
Fish fins as the tissue samples were usually used to extract genomlc DNA in molecular biology studies. In this paper, we introduced a drying method, a convenient method to preserve the fish fins for genomic DNA extraction. This method was applied in common carp (Cyprinus carpio), Amur sturgeon (Acipenser schrenckii), and rainbow trout (Oncorhynchus mykiss ) for genomic DNA extraction. Fin samples were collected, dehydrated by high absorbent paper such as filer paper, and covered with another piece of filter paper, naturally drying. The fins were prevented from moisture and mildew and stored in a cool dry place at room temperature for 3 months. Genomic DNA was extracted using routine phenol-chloroform method. The purity of genomic DNA was measured by ultra- violet spectrophotometer, agarose gel electrophoresis, and polymerase chain reaction (PCR), respectively. The results showed that the OD260/OD280 was varied from 1.82 to 1.89, and that the OD26o/OD:30 from 2.29 to 2.70. The clear and bright electrophoresis bands were observed, and the genomic DNA isolated from fins was suitable for molecular biology experiments. The findings revealed that the dry- ing preservation of fins was feasible to extract high quality genomic DNA as an effective method for precious samples for long term preservation, and was possible and convenient for collection and preservation of massive fin samples of fishes in remote area.
出处
《水产学杂志》
CAS
2014年第1期22-24,共3页
Chinese Journal of Fisheries
基金
国家高技术研究发展计划(2011AA00402-5)
国家科技基础条件平台建设项目
关键词
鱼类
鳍条
保存方法
fish
fin
preservation method