中文摘要：

实时荧光定量PCR技术通过对PCR每一循环扩增产物的实时检测,可对模板的精确拷贝数进行绝对定量,从而用于古DNA实验中提取和扩增条件的比较和优化.本研究采用异硫氰酸胍碱裂解-SiO2吸附的方法,从采自黑龙江省的晚更新世斑鬣狗化石材料中提取得到了斑鬣狗线粒体基因组古DNA.经常规PCR扩增后,将纯化的扩增产物克隆到微生物体内使其大量复制,再用M13通用引物扩增出含少量外源DNA的古DNA目标片段,从而建立了适用于古DNA荧光定量PCR扩增的标准品的制备方法.经检测分析,运用该方法制备的标准品性质稳定,能够准确地指示反应体系中较为精确的古DNA模板拷贝数,从而反映古DNA的提取和扩增效率,用于比较并优化古DNA提取和扩增条件.

英文摘要：

The preparation of the standard samples was essential in ancient DNA amplification.Real time PCR technology was applied to optimize and compare the extraction and amplification of ancient DNAs by calculating the exact copy number of the template.The ancient DNAs were retrieved by guanidinium isothiocyanate lysing and SiO2 absorption from the Late Pleistocene Crocuta crocuta teeth fossils collected at Heilongjiang Province,northeast of China.The partial ancient DNA sequences of the mitochondrial genome D-loop region were PCR amplified,cloned and transfected into E.coli.The prepared standards can be detected with the M13 universal primers in real time PCR.The standard curve and regression analysis showed that the standard sample could be used as monitor of efficiency of extraction and amplification in ancient DNA experiment.