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

摘要

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 gene-modified 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.%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。