定量聚合酶链式反应研究新疆小河古代样本的DNA  

Quantification Polymerase Chain Reaction Designs to Analyze the Ancient Deoxyribonucleic Acid of Xiaohe Cemetery,Xinjiang

作　　者：谢承志[1] 崔银秋[1] 李春香[2] 蔡大伟[1] 王海晶[2] 朱泓[1] 周慧[1] 

机构地区：[1]吉林大学边疆考古研究中心,长春130012 [2]吉林大学生命科学学院,长春130023

出　　处：《分析化学》2007年第5期658-662,共5页Chinese Journal of Analytical Chemistry

基　　金：国家基础科学人才培养基金资助项目(No.J0530184)

摘　　要：利用定量聚合酶链式反应来检测PCR起始阶段的古DNA模板数是鉴定古DNA保存状况以及确保其真实性的有效方法之一。采用SYBRgreenI法对距今3800年左右的新疆小河墓地出土的5例样本线粒体DNA的3个不同长度片段(138bp、209bp和363bp)及核DNA(AMG基因)进行定量。线粒体DNA定量结果显示:小河样本古DNA扩增效率与扩增序列长度成负相关性,只有138bp的结果高于每微升150拷贝。虽然大部分小河样本的AMG基因定量结果都在30拷贝以内,仍然可以进行核DNA的STR或SNP分析。本实验首次通过定量PCR的方法,证明在同一个体中的牙齿中的DNA拷贝数要高于骨骼中的。Human DNA quantification has became an essential analysis to ensure the preservation of ancient samples and the authentication of the result in ancient DNA research. Synergy brands （SYBR） green Ⅰ Realtime PCR was used to quantify five samples which were excavated in Xiaohe cemetery by using the SYBR GREEN PCR Master Mix in Gene Amp 5700 Sequence Detector. Three different fragment sizes （138 bp, 209 bp and 363 bp） within the hypervariable region Ⅰ （ HV1 ） of the mitochondiral deoxyribonuclear acid （mtDNA） and the amelogenin （AMG） gene （121/115 bp） of the nuclear DNA were analyzed. The result showed an inverse correlation between amplicon length and amplification efficiency, only the results of 138 bp were all higher than 150 copies/μL. Although most results of the AMG gene were lower than 30 copies/μL, short tandem repeat （STR） and single nucleotide polymorphism （SNP） of nuclear DNA could be analyzed too. The research also showed that the teeth samples had more DNA copies than bone samples.

关 键 词：定量聚合酶链式反应 古代脱氧核糖核酸 小河墓地 

分 类 号：Q523-3[生物学—生物化学]