Kim Kijeong, Kim Kyung-Yong, Jeon Eunhee, Togloom Ariunaa, Cho Youn-Ock, Lee Min-Soo, Lkhagvasuren Gavaachimed, Choi Jee-Hye, Tumen Dashtseveg, Ja Park Ae, Kim Keun-Cheol, Park Ki-Won, Kim Jae-Hyun, Noh Maengseok, Yoo Kwon-Jong, Lee Kwang-Ho
Institute for Medical Sciences, Chung-Ang University, Seoul 156-756, South Korea.
Am J Phys Anthropol. 2008 May;136(1):114-21. doi: 10.1002/ajpa.20782.
A novel method of ancient DNA (aDNA) purification was developed using ion-exchange columns to improve PCR-amplifiable DNA extraction from ancient bone samples. Thirteen PCR-resistant ancient bone samples aged 500-3,300 years were tested to extract aDNA using a recently reported, silica-based aDNA extraction method and an ion-exchange column method for the further purification. The PCR success rates of the aDNA extracts were evaluated for the amplification ability of the fragments of mitochondrial DNA, a high-copy DNA, and amelogenin, a low-copy DNA. The results demonstrate that the further purification of silica-based aDNA extracts using ion-exchange columns considerably improved PCR amplification. We suggest that the ion-exchange column-based method will be useful for the improvement of PCR-amplifiable aDNA extraction, particularly from the poorly preserved, PCR-resistant, ancient samples.
开发了一种使用离子交换柱的古代DNA（aDNA）纯化新方法，以改进从古代骨骼样本中提取可用于PCR扩增的DNA。使用最近报道的基于硅胶的aDNA提取方法和离子交换柱方法对13个年龄在500至3300年的抗PCR古代骨骼样本进行测试，以提取aDNA并进行进一步纯化。针对线粒体DNA（一种高拷贝DNA）片段和牙釉蛋白（一种低拷贝DNA）的扩增能力，评估了aDNA提取物的PCR成功率。结果表明，使用离子交换柱对基于硅胶的aDNA提取物进行进一步纯化可显著提高PCR扩增效果。我们认为，基于离子交换柱的方法将有助于改进可用于PCR扩增的aDNA提取，特别是从保存不佳、抗PCR的古代样本中提取。
