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A new miniSTR heptaplex system for genetic fingerprinting of ancient DNA from archaeological human bone

机译:一种新的miniSTR七链复合体系统,用于考古人类骨骼中古代DNA的基因指纹

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The analysis of short tandem repeats (STRs) is a useful tool in various contexts of ancient DNA research. Main applications are the reconstruction of kinship, identification, and authentication. Here we describe a short amplicon autosomal short tandem repeat (miniSTR) heptaplex system for the amplification of D13S317, D21S11, D18S51, TH01, D5S818, FGA and Amelogenin from highly degraded DNA as an inexpensive alternative to commercially available kits. All primers were newly designed and the amplicon length of all systems is less than 200 bp, with the exception of some rare alleles in the FGA and D21S11 systems. To validate the suitability of this system for typing STRs from human specimens with low DNA preservation we systematically tested it on 20 skeletal samples from four archaeological sites representing different burial environments and time spans since death. Finally, to test the sensitivity of the heptaplex system, we analyzed serial dilutions of control DNA and ancient DNA extracts. Using the system we were able to reproducibly obtain full STR profiles, down to a concentration of 0.06 ng DNA. Even with 0.004 ng DNA partial profiles could be amplified. The accumulated power of discrimination for the six selected STR loci is 0.99999984, plus the option of genetic sex determination through Amelogenin. The tests conducted prove that the system presented is efficient and especially suited for cases where STRs have to be typed and sex has to be assessed from human specimens with highly degraded DNA.
机译:在古代DNA研究的各种情况下,短串联重复序列(STR)的分析是一种有用的工具。主要应用是亲属关系,身份和身份验证的重建。在这里,我们描述了从高度降解的DNA扩增D13S317,D21S11,D18S51,TH01,D5S818,FGA和Amelogenin的短扩增子常染色体短串联重复(miniSTR)heptaplex系统,作为市售试剂盒的廉价替代品。所有引物都是新设计的,除了FGA和D21S11系统中的一些罕见等位基因外,所有系统的扩增子长度均小于200 bp。为了验证该系统是否适用于从DNA保留率低的人类标本中键入STR,我们在来自四个考古现场的20个骨骼样本上进行了系统测试,这些样本代表了不同的埋葬环境和死后的时间跨度。最后,为了测试七联系统的敏感性,我们分析了对照DNA和古代DNA提取物的系列稀释液。使用该系统,我们能够可再现地获得低至0.06 ng DNA浓度的完整STR谱图。即使使用0.004 ng DNA,也可以扩增部分图谱。六个选定的STR基因座的累计判别力为0.99999984,加上通过Amelogenin进行遗传性别确定的选项。进行的测试证明,所提供的系统是有效的,并且特别适用于必须键入STR且必须从具有高度降解的DNA的人类标本中评估性别的情况。

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