Emulsion PCR-coupled target enrichment: An effective fishing method for high-throughput sequencing of poorly preserved ancient DNA
Emulsion PCR-coupled target enrichment: An effective fishing method for high-throughput sequencing of poorly preserved ancient DNA
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乳液 PCR 耦合靶标富集：一种对保存不良的古 DNA 进行高通量测序的有效捕获方法
DOI：
10.1016/j.gene.2013.07.040
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发表时间：
2013-10-10
期刊：
影响因子：
3.5
通讯作者：
Ueda, Shintaroh
中科院分区：
文献类型：
--
作者：
Kihana, Makio;Mizuno, Fuzuki;Ueda, Shintaroh
由于深度测序的困难，古DNA的高通量测序一直局限于保存异常完好的古代材料。主要原因是埋藏物中普遍存在的微生物侵袭，导致提取的DNA中微生物DNA的相对比例急剧增加。我们提出了一个统一的策略，其中乳液PCR加上目标富集，然后进行下一代测序。该方法使得有可能获得映射到感兴趣的靶序列的有效的非重复读段，并且这可以实现来自典型材料的古老DNA的深度和可靠的测序，即使保存较差。(C)2013爱思唯尔有限公司版权所有。
Due to the difficulties in deep sequencing, high-throughput sequencing of ancient DNA has been limited to exceptionally well-preserved ancient materials. The primary factor is microbial attack popularly observed lit the buried materials, and it causes drastic increase in relative ratio of microbial DNA in the extracted DNA. We present a unified strategy in which emulsion PCR is coupled with target enrichment followed by next-generation sequencing. The method made it possible to obtain efficiently non-duplicated reads mapped to target sequences of interest, and this can achieve deep and reliable sequencing of ancient DNA from typical materials, even though poorly preserved. (C) 2013 Elsevier B.V. All rights reserved.