Evaluation of whole genome amplification methods using postmortem brain samples
Evaluation of whole genome amplification methods using postmortem brain samples
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DOI：
10.1016/j.jneumeth.2007.05.029
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发表时间：
2007-09-15
影响因子：
3
通讯作者：
Kato, Tadafumi
中科院分区：
文献类型：
--
作者：
Iwamoto, Kazuya;Ueda, Junko;Kato, Tadafumi
最近的发现强调了检查源自人脑的基因组 DNA 的重要性，例如 DNA 甲基化与行为或精神障碍之间可能存在的联系，以及来自同一个体的神经元之间可能由转座子和非整倍体引起的基因组差异。因此，获得足够量的人脑基因组DNA是进一步研究的关键问题。全基因组扩增 (WGA) 方法通常以 10(4)-10(6) 的量级扩增基因组 DNA，将成为为各种分子遗传学研究提供足够量 DNA 的宝贵工具。在这里，我们评估了三种方法，包括基于 PCR 和非 PCR 的 WGA，以及使用冷冻死后脑组织的 DNA 提取方法。我们发现，如果使用特定的 DNA 提取方案，来自死后大脑的 WGA 产品可用于分子遗传分析，并且最合适的 WGA 方法取决于要扩增的基因组 DNA 的状态。 (c) 2007 Elsevier B.V. 保留所有权利。
The importance of examining genomic DNA derived from human brain has been highlighted by recent findings such as the possible link between DNA methylation and behavior or mental disorders, as well as the possible genomic differences between neurons from the same individual caused by transposons and aneuploidy. Consequently, obtaining a sufficient amount of genomic DNA derived from human brain is a critical issue for further research. Whole genome amplification (WGA) methods, by which genomic DNA is typically amplified on the order of 10(4)-10(6), will be a valuable tool for providing a sufficient amount of DNA for various molecular genetic studies. Here we evaluated three methods, including both PCR-based and non-PCR based WGA, as well as DNA extraction methods using frozen postmortem brain tissue. We found that WGA products from postmortem brains can be used in molecular genetic analysis, if a particular protocol for DNA extraction is used, and the most appropriate method for WGA depends on the state of the genomic DNA to be amplified. (c) 2007 Elsevier B.V. All rights reserved.