Reyes Aurelio, Yasukawa Takehiro, Holt Ian J
MRC Dunn Human Nutrition Unit, Wellcome Trust, Cambridge, United Kingdom.
Methods Mol Biol. 2007;372:219-32. doi: 10.1007/978-1-59745-365-3_16.
Replication intermediates can be separated on agarose gels in two dimensions to reveal a wealth of data on mechanisms of DNA replication. When applied to mitochondrial DNA of higher vertebrates, this technique unearthed a host of unexpected findings, the full implications of which are still being absorbed. Here, we describe the procedures we use to isolate intact mitochondrial replication intermediates from liver of higher vertebrates and the process of separating DNA fragments on neutral two-dimensional agarose gels.
复制中间体可以在琼脂糖凝胶上进行二维分离，以揭示大量有关DNA复制机制的数据。当应用于高等脊椎动物的线粒体DNA时，这项技术发现了许多意想不到的结果，其全部意义仍在被人们理解。在这里，我们描述了从高等脊椎动物肝脏中分离完整线粒体复制中间体的程序，以及在中性二维琼脂糖凝胶上分离DNA片段的过程。
