模式小鼠批量鉴定方法的探讨  

Batch identification Methods for model mice

作　　者：金悠 倪岚[1] 魏琦 赵欢 何金钢 JIN You;NI Lan;WEI Qi;ZHAO Huan;HE Jingang(National Demonstration Center for Experimental Basic Medical Education,Huazhong University of Science and Technology,Wuhan 430030,China;Department of Pharmacology,School of Basic Medicine,Tongji MedicalCollege,Huazhong University of Science and Technology,Wuhan 430030)

机构地区：[1]华中科技大学基础医学国家级实验教学示范中心,武汉430030 [2]华中科技大学同济医学院基础医学院药理学系,武汉430030

出　　处：《中国比较医学杂志》2025年第1期86-91,共6页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金面上项目(82273911)。

摘　　要：目的为模式小鼠基因型批量鉴定工作建立高效的DNA提取方法。方法以鼠尾组织为实验材料,采用碱煮简化法、碱煮常规法、蛋白酶K裂解法和DNA提取试剂盒4种方法提取模式小鼠DNA,测定DNA模板纯度、浓度,评价凝胶电泳图的优劣,并比较4种方法所耗时长及实验成本。结果蛋白酶K裂解法提取的DNA浓度最高,碱煮简化法和常规法次之;试剂盒处理所得的DNA纯度较高,碱煮简化法次之。4种方法处理所得的DNA模板通过聚合酶链反应(polymerase chain reaction,PCR)扩增、凝胶电泳后均可得到清晰的DNA目的条带。此外,碱煮简化法提取的DNA模板在-20℃保存1个月后仍可用于基因鉴定,且耗时最少、成本最低。结论模式小鼠批量鉴定基因型时,碱煮简化法是目前最简便、最快捷、最经济的DNA模板提取方法。Objective To establish an efficient DNA extraction method for the batch identification of genotypes in model mice.Methods We extracted total DNA from transgenic mouse tails using four Methods:alkaline simplified group,alkaline routine group,protease K cleavage group and DNA extraction kit group.The purity and concentration of DNA obtained by the four Methods were measured,the effects of gel electrophoresis were evaluated,and the time and experimental costs of the four Methods were compared.Results The protease K cleavage method produced the highest concentration of DNA,followed by the simplified alkaline boiling and routine alkaline boiling method.The reagent kit produced the highest DNA purity,followed by the simplified alkaline boiling method.The DNA templates obtained by the four method could be amplified by polymerase chain reaction and gel electrophoresis to obtain clear DNA target bands.In addition,the DNA template extracted by the simplified alkaline boiling method could be used for gene identification after storage at-20℃for 1 month,as well as requiring the least time and lowest costs.Conclusions The simplified alkaline boiling method is currently the simplest,fastest,and most economical DNA template-extraction method for batch identification of genotypes in model mice.

关 键 词：基因型鉴定 DNA提取 聚合酶链反应 模式小鼠 

分 类 号：R-33[医药卫生]