湖北医药学院学报2011，Vol.30Issue(6)：557-560,4.
应用降落PCR及克隆测序检测2型糖尿病外周血白细胞胰岛素样生长因子受体IGF1R基因甲基化
Applying Touchdown-PCR and Clone-based Sequencing to Detect the Methylation of IGF1R Gene Promoter in the Peripheral Leukocyte of Type 2 Diabetes Mellitus
摘要
Abstract
Objective To apply touchdown-PCR and clone-based sequencing detecting the methylation sites of IGF1R gene promoter in the peripheral leukocyte of type 2 diabetes mellitus. Methods After the bisulfite treatment of the DNA extracted from the peripheral leukocyte in patients with type 2 diabetes mellitus, the primer was design to amplify the target fragments by general PCR and touchdown-PCR,then the target fragments were used to direct sequencing and clone sequencing. Re-SUltS Compared the amplify results betweem touchdown-PCR and general PCR,the former had clearer fragment,more products, less dimers and saved time to explore the annealing temperature. The results of clone sequencing displayed clearly and accurately of every base. Conclusion Touchdown-PCR were recommend to be used to detect the methylation status after the DNA underwent the bisulfite treatment, the clone sequencing is a advisable choice.关键词
甲基化/降落PCR/胰岛素样生长因子1受体/2型糖尿病/测序Key words
Methylation/Touchdown-PCR/Insulin-like growth factor-1 receptor/Type 2 diabetes mellitus/Sequence分类
医药卫生引用本文复制引用
陈玲,严世荣,胡培,陈永梅,程龙海..应用降落PCR及克隆测序检测2型糖尿病外周血白细胞胰岛素样生长因子受体IGF1R基因甲基化[J].湖北医药学院学报,2011,30(6):557-560,4.基金项目
湖北医药学院博士启动金(2010QDJ22) （2010QDJ22）
