Iwamoto Kazuya, Ueda Junko, Nakano Yoko, Bundo Miki, Ukai Wataru, Hashimoto Eri, Saito Toshikazu, Kato Tadafumi
Laboratory for Molecular Dynamics of Mental Disorders, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
J Neurosci Methods. 2007 Sep 15;165(1):104-10. doi: 10.1016/j.jneumeth.2007.05.029. Epub 2007 Jun 2.
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.
近期的研究发现凸显了检测源自人类大脑的基因组DNA的重要性，这些发现包括DNA甲基化与行为或精神障碍之间可能存在的联系，以及转座子和非整倍体导致的同一个体神经元之间可能存在的基因组差异。因此，获取足够量的源自人类大脑的基因组DNA是进一步研究的关键问题。全基因组扩增（WGA）方法通常可将基因组DNA扩增10⁴至10⁶倍，将成为为各种分子遗传学研究提供足够量DNA的宝贵工具。在此，我们评估了三种方法，包括基于PCR和非基于PCR的WGA方法，以及使用冷冻尸检脑组织的DNA提取方法。我们发现，如果使用特定的DNA提取方案，来自尸检大脑的WGA产物可用于分子遗传学分析，并且WGA的最合适方法取决于待扩增基因组DNA的状态。
