“逆向根管技术”提取古人类牙齿DNA的初步探索

"Reverse-root-canal technique" to extract DNA from ancient teeth

目的:建立一种利用古人类牙齿提取基因组DNA的简便方法。方法:选取西安地区出土的3 000年前人牙7个、1 000年前人牙1个,现代人牙2个;从根尖末端开口分离根管内样本,结合Chelex-100基因组DNA提取法提取牙内部组织的DNA;设计釉原蛋白基因特异性引物,采用PCR技术进行样本的性别鉴定。结果:所选样本基因组DNA分子含量较高,可用于PCR扩增技术相关的性别鉴定,经鉴定本研究中的10个样本为1个男性9个女性。结论:"逆向根管技术"与Chelex-100提取法相结合具有低损伤的特征,能够较大程度地保存古人类牙齿外形,并且操作简便,可用于考古、遗传等多学科的研究。

AIM： To find a convenient method for DNA extraction from ancient teeth.METHODS： Eight unearthed teeth in the area of Xi＇an were chosen in this study,one of which was about 1000 years ago and the others were 3000 years ago.Two teeth from modern people were chosen as the control.The ＂Reverse-root-canal technique＂ and Chelex-100 were applied to extract DNA from the teeth samples.The specific primers of amelogenin were designed and PCR method was used to perform the sex typing.RESULTS： The genomic DNA extracted from the samples had a high quality for PCR amplication and sex determination.Of the ten samples in the study,one was male and nine were female.CONCLUSION： The ＂Reverse-root-canal technique＂ is a low-destructive method for obtaining DNA from ancient teeth.It is convenient for research in archaeology,genetics and other science branches.