Department of Medical Biochemistry and Biophysics, Umeå University, 901 87 Umeå, Sweden.
Department of Medical Biochemistry and Biophysics, Umeå University, 901 87 Umeå, Sweden.
Mitochondrion. 2021 Nov;61:179-187. doi: 10.1016/j.mito.2021.10.005. Epub 2021 Oct 30.
The integrity of mitochondrial DNA (mtDNA) isolated from solid tissues is critical for analyses such as long-range PCR, but is typically assessed under conditions that fail to provide information on the individual mtDNA strands. Using denaturing gel electrophoresis, we show that commonly-used isolation procedures generate mtDNA containing several single-strand breaks per strand. Through systematic comparison of DNA isolation methods, we identify a procedure yielding the highest integrity of mtDNA that we demonstrate displays improved performance in downstream assays. Our results highlight the importance of isolation method choice, and serve as a resource to researchers requiring high-quality mtDNA from solid tissues.
从固体组织中分离出的线粒体 DNA（mtDNA）的完整性对于长距离 PCR 等分析至关重要，但通常在无法提供个体 mtDNA 链信息的条件下进行评估。我们使用变性凝胶电泳表明，常用的分离程序会产生每条链含有几个单链断裂的 mtDNA。通过对 DNA 分离方法的系统比较，我们确定了一种产生 mtDNA 完整性最高的方法，我们证明该方法在下游检测中具有更好的性能。我们的研究结果强调了分离方法选择的重要性，并为需要从固体组织中获得高质量 mtDNA 的研究人员提供了资源。
