基于牙釉质基因巢式PCR性别鉴定超微量DNA检测方法的建立  

Ultratrace DNA Detection Based on PCR Amplification Amelogenin Gene Associated with Sex Identification

作　　者：唐纪伟[1] 高庆华[1] 

机构地区：[1]塔里木大学畜牧科技兵团重点实验室,新疆阿拉尔843300

出　　处：《塔里木大学学报》2011年第2期24-27,共4页Journal of Tarim University

基　　金：国家自然科学基金项目(30760170);国家科技支撑计划课题子课题(2008BADB2B05-1-3)

摘　　要：实验根据山羊牙釉质(AMEL)基因序列设计巢式引物,用紫外分光仪检测基因组DNA的浓度,梯度稀释(10-2μg/μL、10-3μg/μL,10-4μg/μL,10-5μg/μL),巢式PCR扩增后琼脂糖凝胶电泳检测。结果表明,利用巢式PCR方法对10 pg量基因组DNA(约3个细胞)进行扩增,得到明显目的条带。巢式PCR扩增AMEL基因鉴定性别的方法具有较高的准确性和敏感性,超微量DNA检测方法对胚胎移植前性别鉴定及提高切割胚胎移植的成活率方面具有重要的意义。In this experiment,hircine blood genomic DNA was extracted,nested primer was designed by AMEL gene of goat,and the concentration of the DNA was tested by Ultraviolet spectrophotometer,then blood genomic DNA of goats was serially diluted（10-2μg/μL,10-3 μg/μL,10-4μg/μL,10-5 μg/μL）,and nested PCR amplification was tested by Agarose gel electrophoresis.The results show that the method of nested PCR using 10 pg genomic DNA（approximately 3 cells）,could get clear purpose band.The method based on sex identification by nesred PCR amplifying amelogenin gene was highly accurate and sensitive.There is an important significance about sex identification before embryo transfer and enhancing the survival rates of cutting embryo transplantation by using ultratrace DNA detection.

关 键 词：巢式PCR 超微量DNA 牙釉质基因1 

分 类 号：S852.65[农业科学—基础兽医学]