碱性裂解液提取DNA法在环氧化酶亚型基因敲除小鼠基因型鉴定中的应用  

Application of a DNA Extraction Method Using Alkaline Lysis Buffer in the Genotyping of COX Isoform Knock-out Mice

作　　者：李振华[1] 张英展[1] 周应毕[1] 刘斌[1] 

机构地区：[1]汕头大学医学院,广东汕头515041

出　　处：《中国医学创新》2014年第6期6-8,共3页Medical Innovation of China

基　　金：国家自然科学基金资助项目(81000111)

摘　　要：目的:应用碱性裂解液法提取的DNA鉴定环氧化酶(COX)亚型基因敲除小鼠,为鉴定纯合子小鼠提供一种方便快捷、低成本、可靠的实验方法。方法:取子代小鼠耳缘组织于EP管中,用碱性裂解液提取基因组DNA,PCR扩增COX-1和COX-2基因片段,通过琼脂糖凝胶电泳对小鼠基因型作出鉴定。结果:碱性裂解液法提取的DNA能应用于子代小鼠的不同基因型的鉴定,即野生型(COX-1+/+;COX-2+/+)、杂合子(COX-1+/-;COX-2+/-)和纯合子(COX-1-/-;COX-2-/-)。结论:建立了碱性裂解液提取DNA法,并将该法成功应用于环氧化酶亚型基因敲除小鼠基因型鉴定中。Objective：To identify Cyclo-oxygenase（COX）isoform knock-out mice with DNA extracted with alkaline lysis buffer and thus to provide a convenient,low-cost and reliable method to identify the homozygous mice.Method：Genome DNA was extracted from the marginal ear tissue of filial generation mice by alkaline lysis buffer and PCR was employed to amplify gene fragments of COX-1 or COX-2,then the genotyping of mice was performed by agarose gel electrophoresis.Result：DNA was conveniently extracted with the method,and with the DNA,genotypes,namely wild-type（COX-1＋/＋,COX-2＋/＋）,heterozygote（COX-1＋/-,COX-2＋/-） and homozygote（COX-1-/-,COX-2-/-）,of the offsprings of COX-1 or COX-2 genemodified mice were reliably identified.Conclusion：A DNA extraction method using alkaline lysis buffer has been established and can be successfully applied to the genotyping of COX isoforms knock-out mice.

关 键 词：碱性裂解液 环氧化酶 基因敲除小鼠 

分 类 号：R346[医药卫生—基础医学]