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模式小鼠总DNA三种提取方法比较
引用本文:刘甦苏,左琴,周舒雅,王辰飞,贺争鸣,李保文,范昌发. 模式小鼠总DNA三种提取方法比较[J]. 中国实验动物学杂志, 2014, 0(7): 45-50
作者姓名:刘甦苏  左琴  周舒雅  王辰飞  贺争鸣  李保文  范昌发
作者单位:中国食品药品检定研究院,国家啮齿类实验动物种子中心,北京100050
基金项目:中国食品药品检定研究院中青年发展研究基因(2012B5).
摘    要:目的:建立简便、快捷、经济的模式小鼠总DNA提取方法,以快速鉴定大批量模式小鼠基因型。方法采用苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法提取同种模式小鼠总DNA,对比DNA纯度、得率、耗费时间,并比较基因型鉴定结果。结果苯酚抽提法得率最高,异丙醇沉淀法最低;而纯度则按照苯酚抽提法、异丙醇沉淀法、鼠耳煮沸法顺序递减;在耗时上鼠耳煮沸法最短。三种方法提取的DNA均可做模版用于基因型鉴定。结论鼠耳煮沸法操作简单、成本最低,快速、基因型鉴定结果可靠,可用于规模化的基因型鉴定实验中。

关 键 词:总DNA提取方法  基因型鉴定  PCR反应  模式小鼠

Comparison of three methods for total DNA extraction from mouse models
LIU Su-su,ZUO Qin,ZHOU Shu-ya,WANG Chen-fei,HE Zheng-ming,LI Bao-wen,FAN Chang-fa. Comparison of three methods for total DNA extraction from mouse models[J]. Chinese Journal of Laboratory Animal Science, 2014, 0(7): 45-50
Authors:LIU Su-su  ZUO Qin  ZHOU Shu-ya  WANG Chen-fei  HE Zheng-ming  LI Bao-wen  FAN Chang-fa
Affiliation:(Laboratory Animal Resource Center, National Institute of Food and Drug Control, Beijing 100050, China)
Abstract:Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P 〈0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .
Keywords:Total DNA extraction  Genotype identification  PCR reaction  Model mice
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