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 |
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引用本文: | 李振华,张英展,周应毕,刘斌. 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[J]. 现代保健, 2014, 0(6): 6-8 |
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作者姓名: | 李振华 张英展 周应毕 刘斌 |
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作者单位: | 汕头大学医学院,广东汕头515041 |
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基金项目: | 国家自然科学基金资助项目(81000111) |
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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。
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关 键 词: | 碱性裂解液 环氧化酶 基因敲除小鼠 |
Application of a DNA Extraction Method Using Alkaline Lysis Buffer in the Genotyping of COX Isoform Knock-out Mice |
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Affiliation: | LI Zhen-hua;ZHANG Ying-zhan;ZHOU Ying-bi( Shantou University Medical College, Shanu 515041, China) |
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Abstract: | 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. |
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Keywords: | Alkaline lysis buffer Cyclo-oxygenase Gene knockout mouse |
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