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次氯酸钠去污处理对从人牙中回收DNA的影响。

Effect of sodium hypochlorite decontamination on the DNA recovery from human teeth.

作者信息

Koehn Katharina, Buettner Andreas, Lindner Iris

机构信息

Institute of Legal Medicine, Rostock University Medical Center, St.-Georg-Str. 108, 18055, Rostock, Germany.

出版信息

Int J Legal Med. 2020 Jan;134(1):93-99. doi: 10.1007/s00414-019-02174-2. Epub 2019 Nov 6.

Abstract

Genetic identification of skeletal human remains is often realized by short tandem repeat (STR) genotyping of nuclear DNA. Dental DNA is preferred to DNA from bone for the better protection of the endogenous DNA. Especially if whole tooth grinding is intended to access the DNA, contaminations with exogenous DNA have to be avoided. The immersion of the tooth in sodium hypochlorite (NaOCl, known as bleach) is one common procedure to clean the outer surface from extraneous DNA and PCR inhibitors. To investigate the impact of bleaching on endogenous DNA and the decontamination success, 71 recently extracted teeth were differently treated with sodium hypochlorite (2.5 or 5.0% NaOCl for 30 or 60 s, 5.0% NaOCl for 10 min, and control group) in the beginning of the extraction process, whereas equally handled afterwards. Quantitative and qualitative evaluation of the extracted DNA was performed. There was a great variation for the DNA concentration of the extracts even within a group of the same NaOCl treatment. Complete DNA profiles from single persons with alleles for the 16 ESS (European Standard Set) STR loci were obtained for all regarded teeth. A statistically significant difference between the DNA yields of the treatment groups was not determined. Moreover, a negative effect of NaOCl (2.5% and 5.0%) on the DNA recovery could not be observed. Significant larger amounts of DNA were extracted from anterior teeth in contrast to posterior teeth.

摘要

人类骨骼遗骸的基因鉴定通常通过对核DNA进行短串联重复序列(STR)基因分型来实现。由于牙源性DNA能更好地保护内源性DNA,因此相较于骨源性DNA,牙源性DNA更受青睐。特别是在打算通过全牙研磨获取DNA时,必须避免外源性DNA的污染。将牙齿浸入次氯酸钠(NaOCl,即漂白剂)中是一种常见的从外源性DNA和PCR抑制剂中清洁牙齿外表面的方法。为了研究漂白对内源性DNA的影响以及去污效果,在提取过程开始时,对71颗最近提取的牙齿用次氯酸钠进行了不同处理(2.5%或5.0%的NaOCl处理30或60秒、5.0%的NaOCl处理10分钟,以及对照组),之后进行相同处理。对提取的DNA进行了定量和定性评估。即使在同一NaOCl处理组内,提取物的DNA浓度也存在很大差异。对于所有研究的牙齿,均获得了来自单个人的16个ESS(欧洲标准集)STR基因座等位基因的完整DNA图谱。未确定各处理组DNA产量之间存在统计学上的显著差异。此外,未观察到NaOCl(2.5%和5.0%)对DNA回收率有负面影响。与后牙相比,从恒牙中提取的DNA量明显更多。

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