介绍一种快速提取胎鼠基因组DNA进行基因型分析的改良方法  

Rapid Preparation of High Quality Genomic DNA from Mouse Embryo for Genotyping

作　　者：杨瑞[1] 麻献华[1] 唐君仪[2] 章卫平[1] 谢志芳[3] 

机构地区：[1]第二军医大学基础部病理生理学教研室,上海200433 [2]第二军医大学研究生管理大队学员二队,200433 [3]第二军医大学细胞生物学教研室,上海200433

出　　处：《医学研究杂志》2013年第5期46-48,共3页Journal of Medical Research

基　　金：国家自然科学基金资助项目(31270814)

摘　　要：目的建立一种快速提取胎鼠基因组DNA进行基因型分析的简便方法。方法先对胎鼠组织进行碱裂解,然后通过盐析纯化DNA,经琼脂糖凝胶电泳判断DNA大小,检测A260/A280的比值判断其纯度,最后将抽提的DNA进行PCR扩增和基因型分析,检测其作为PCR模板的稳定性。结果在2h内能分离到胎鼠基因组DNA,片段完整没有明显的降解现象,A260/A280比值>1.77,能稳定用于PCR基因型鉴定分析。结论该方法简便、快速、经济,能得到PCR级的高质量胎鼠基因组DNA,为准确和快速地进行胎鼠的基因型分析提供保证,具有很好的稳定性、可靠性和实用性。Objective To develop a rapid method for the extraction of high quality genomic DNA from mouse embryo for genotyping. Methods Genomic DNA was prepared using alkaline lysis followed by high - salt extraction from mouse embryonic tissue. The integrity and purity of the DNA was assessed by performing agarose gel electrophoresis and ratio absorbance measurements at A260/A280. The DNA samples were then subjected to PCR amplification using two primer pairs. Results The genomic DNA was extracted from mouse embryonic tissue within 2h using the new method. There was no sign of degraded DNA during preparation and the purity of the DNA determined from the A260/A280 ratio averaged 〉 1.77. The DNA produced a reproducible PCR product pattern. Conclusion The new method is time - saving, economic and reproducible for the preparation of high - quality genomic DNA from mouse embryo for genotyping.

关 键 词：基因组 DNA 分离 胎鼠 PCR 基因型鉴定 

分 类 号：R392-33[医药卫生—免疫学]