Strehle Mackenzie M, Purfeerst Emma, Christensen Alan C
School of Biological Sciences, University of Nebraska, Lincoln, NE, USA.
Mitochondrial DNA B Resour. 2018 Feb 15;3(1):239-242. doi: 10.1080/23802359.2018.1438856.
Current mitochondrial purification techniques are tedious and protracted due to their emphasis on recovering physiologically active mitochondria. However, for studies that are exclusively interested in isolating mitochondrial DNA (mtDNA) for applications such as PCR and sequencing, respiring mitochondria - and the complex procedures that stem from the need to retain their function - are unnecessary. Still, global DNA extraction methods have proven insufficient for mitochondrial DNA isolation because nuclear mitochondrial DNA segments (NUMTs) pose unique challenges to accurate mtDNA quantification and characterization. We present a rapid and simple extraction technique that maximizes recovery of mitochondrial DNA from plant cells, while minimizing the presence of nuclear DNA. Through real-time PCR, we show that this method provides a significant increase in the enrichment of mitochondrial DNA compared to that of nuclear DNA in both and . This method has important implications for future mitochondrial DNA analyses as it possesses few procedural limitations and minimizes the analytical problems typically associated with mtDNA purification by other techniques.
当前的线粒体纯化技术既繁琐又耗时，因为这些技术着重于回收具有生理活性的线粒体。然而，对于那些仅旨在分离线粒体DNA（mtDNA）以用于PCR和测序等应用的研究而言，进行呼吸作用的线粒体以及因需要保留其功能而产生的复杂程序是不必要的。尽管如此，已证明常规的DNA提取方法不足以用于线粒体DNA的分离，因为核线粒体DNA片段（NUMTs）给准确的mtDNA定量和表征带来了独特的挑战。我们提出了一种快速简便的提取技术，该技术能最大限度地从植物细胞中回收线粒体DNA，同时尽量减少核DNA的存在。通过实时PCR，我们表明，与[此处原文缺失相关信息]中的核DNA相比，该方法能显著提高线粒体DNA的富集度。此方法对未来的线粒体DNA分析具有重要意义，因为它几乎没有程序上的限制，并且将通常与其他技术进行mtDNA纯化相关的分析问题降至最低。
