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首页> 外文期刊>Journal of Neuroscience Methods >Single extraction protocol for the analysis of 8-hydroxy-2'-deoxyguanosine (oxo8dG) and the associated activity of 8-oxoguanine DNA glycosylase.
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Single extraction protocol for the analysis of 8-hydroxy-2'-deoxyguanosine (oxo8dG) and the associated activity of 8-oxoguanine DNA glycosylase.

机译：用于分析8-hydroxy-2'-deoxyguanosine（oxo8dG）和8-oxoguanine DNA糖基化酶相关活性的单一提取方案。

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摘要

Determination of the promutagenic base 8-hydroxy-2'-deoxyguanosine (oxo(8)dG) has been the hallmark of studies aimed to determine oxidative damage to DNA. Different techniques including HPLC, GC-mass spectrometry, DNA sensitive sites and histology have been used to quantify oxo(8)dG levels in samples from different sources. The most accepted and well-established methods are based on HPLC and the ability of oxo(8)dG to be oxidized with an electrochemical detector. Considerable concerns have been raised in the ability of different labs to utilize a process of DNA extraction that reduces the levels of artifactual oxo(8)dG formed during sample workup. Here, we present a fully detailed protocol that has been extensively used in our Lab to extract and analyze DNA and has little or no impact in the basal levels of oxo(8)dG. Additionally, this protocol allows for the determination of the activity of mOgg1, the enzyme responsible for the initial step in the repair of the accumulated oxo(8)dG, in the same samplein which oxo(8)dG is detected.
机译：致突变的碱基8-羟基-2'-脱氧鸟苷（oxo（8）dG）的确定已成为旨在确定DNA氧化损伤的研究的标志。包括HPLC，GC-质谱，DNA敏感位点和组织学在内的不同技术已用于量化来自不同来源的样品中oxo（8）dG的水平。最被接受和公认的方法是基于HPLC和oxo（8）dG被电化学检测器氧化的能力。人们对不同实验室利用DNA提取工艺的能力提出了极大的关注，该工艺可降低样品处理过程中形成的人为oxo（8）dG的水平。在这里，我们介绍了一个非常详细的协议，该协议已在我们的实验室中广泛用于提取和分析DNA，并且对oxo（8）dG的基础水平几乎没有影响。此外，此协议允许确定mOgg1的活性，该酶负责在检测到oxo（8）dG的同一样品中修复累积的oxo（8）dG的初始步骤。

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