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.