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A simple and efficient method for PCR amplifiable DNA extraction from ancient bones

机译:一种从古骨中PCR扩增DNA的简单有效方法

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摘要

A simple and effective modified ethanol precipitation-based protocol is described for the preparation of DNA from ancient human bones. This method is fast and requires neither hazardous chemicals nor special devices. After the powdering and incubating of the bone samples Dextran Blue was added as a carrier for removing the PCR inhibitors with selective ethanol precipitation. This method could eliminate the time-consuming separate decalcification step, dialysis, application of centrifugation-driven microconcentrators and the second consecutive PCR amplification. The efficiency of this procedure was demonstrated on ten 500–1200-year-old human bones from four different Hungarian burial sites. A mitochondrial specific primer pair was used to obtain sequence information from the purified ancient DNA. The PCR amplification, after our DNA extraction protocol, was successful from each of the 10 bone samples investigated. The results demonstrate that extraction of DNA from ancient bone samples with this new approach increases the success rate of PCR amplification.
机译:描述了一种简单有效的基于乙醇沉淀的改良方案,用于从古代人类骨骼中制备DNA。此方法快速,不需要有害化学物质或特殊设备。在将骨样品撒粉并孵育后,添加右旋糖酐蓝作为载体,通过选择性乙醇沉淀去除PCR抑制剂。这种方法可以消除耗时的单独脱钙步骤,透析,离心驱动微量浓缩器的应用以及第二次连续PCR扩增。在四个不同的匈牙利埋葬地点的十个500-1200岁的人类骨骼上证明了该程序的效率。线粒体特异性引物对用于从纯化的古代DNA中获得序列信息。经过我们的DNA提取方案后,从所研究的10个骨骼样品中成功扩增了PCR。结果表明,使用这种新方法从古代骨骼样品中提取DNA可以提高PCR扩增的成功率。

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