古人类长骨和髋骨基因组DNA的体外分离及分子表征

Dinesh Kumar, L. Jeena, Devendra Shekhawat, Ira Verma, Sidd Sana
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引用次数: 0

摘要

背景: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提取基因组DNA。基于DNA纯度(260/280)和PCR方法对基因组DNA进行定性和定量评价。方法和材料:本研究共采集50块防腐古骨,包括20块长骨和30块髋骨样本进行基因组DNA提取。基因组DNA提取的效率在50岁以上的古代人类HIP和长骨样本上得到了证明。通过0.8%琼脂糖凝胶电泳和PCR扩增对提取的基因组DNA进行体外定量和定性分析。为了评估提取的基因组DNA的质量,使用线粒体特异性ATPase6基因引物获得675bp的序列信息。结果:我们的数据显示,在260/280时,基因组DNA的浓度在1.6和2.0之间导致成功的PCR扩增。我们的研究结果表明,用手动方法从古代骨骼样本中提取DNA将提高聚合酶链式反应(PCR)的扩增效率。结论:在本研究中,描述了一种简单、省时、成本效益高的从古代人类骨骼中提取基因组DNA的方案。此外,我们相信用这种方法从古代骨骼样本中提取基因组DNA将提高PCR扩增的成功率。
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In Vitro Isolation and Molecular Characterization of Genomic DNA from Ancient Human Long and Hip Bones
Background: A simple and effective modified ethanol precipitation-based protocol was described for the extraction of genomic DNA from ancient human bones. The qualitative and quantitative evaluation of genomic DNA was done based on DNA purity (260/280) and the PCR method. Method and Materials: In this study, a total of 50 embalmed ancient bones, including 20 long and 30 hip bone samples, were taken for genomic DNA extraction. The efficiency of the genomic DNA extraction was demonstrated on >50-year-old ancient human HIP and long bone samples. In vitro quantitative and qualitative analysis of extracted genomic DNA was performed by 0.8% agarose gel electrophoresis and PCR amplification. To assess the quality of extracted genomic DNA, a mitochondrial-specific ATPase6 gene primer was used to obtain sequence information of 675 bp. Result: Our data show that a concentration of genomic DNA between 1.6 and 2.0 at 260/280 resulted in successful PCR amplification. Our results demonstrated that the extraction of DNA from ancient bone samples with a manual approach will increase the amplification efficiency of the polymerase chain reaction (PCR). Conclusion: In the present study, a simple, time-saving, and cost-effective protocol is described for the extraction of genomic DNA from ancient human bones. Further, we believe the extraction of genomic DNA from ancient bone samples with this approach will increase the success rate of PCR amplification.
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