A New Method of the Visualization of the Double-Stranded Mitochondrial and Nuclear DNA
A New Method of the Visualization of the Double-Stranded Mitochondrial and Nuclear DNA
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DOI：
10.1371/journal.pone.0066864
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发表时间：
2013-06-18
期刊：
影响因子：
3.7
通讯作者：
Koberna, Karel
中科院分区：
文献类型：
--
作者：
Ligasova, Anna;Strunin, Dmytro;Koberna, Karel
这项研究描述了一种灵敏的原位检测双链DNA分子的方法。它是基于铜(I)在氧气存在下对脱氧核糖部分的氧化攻击。我们以前已经证明，氧化攻击会导致DNA中频繁的缺口的形成。在这里，我们证明了缺口可以作为DNA聚合酶I有效合成互补标记链的起点，并且这种双链DNA的酶检测是一种敏感的方法，能够在甲醛固定的人类细胞中原位检测核和线粒体基因组。
The study describes the method of a sensitive detection of double-stranded DNA molecules in situ. It is based on the oxidative attack on the deoxyribose moiety by copper(I) in the presence of oxygen. We have shown previously that the oxidative attack leads to the formation of frequent gaps in DNA. Here we have demonstrated that the gaps can be utilized as the origins for an efficient synthesis of complementary labeled strands by DNA polymerase I and that such enzymatic detection of the double-stranded DNA is a sensitive approach enabling in-situ detection of both the nuclear and mitochondrial genomes in formaldehyde-fixed human cells.