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首页> 外文期刊>Forensic science international >Use of bleach to eliminate contaminating DNA from the surface of bones and teeth.
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Use of bleach to eliminate contaminating DNA from the surface of bones and teeth.

机译:使用漂白剂消除骨骼和牙齿表面的污染性DNA。

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摘要

The extraction of DNA from archaeological or forensic skeletal remains can provide quite powerful data for analysis, but is plagued by a unique set of methodological problems. One of the most important methodological problems to overcome in such analyses is the presence of modern contamination on the surfaces of bones and teeth, which can lead to false positives and erroneous results unless it is removed before DNA extraction is initiated. Ancient DNA (aDNA) researchers and forensic scientists have employed a number of techniques to minimize such contamination. One such technique is the use of bleach (sodium hypochlorite-NaOCl) to "destroy" contaminating DNA. However, a consensus on the optimum concentration of sodium hypochlorite to be used and the amount of time the bone or tooth should be exposed to it has not emerged. The present study systematically approaches the issue by introducing contamination to ancient bones (from approximately 500 BP) and determining which of several sodium hypochlorite treatments best eliminates surface contamination. The elimination of surface contamination from bone requires immersion in at least 3.0% (w/v) sodium hypochlorite (approximately equal parts of commercial bleach and water) for at least 15min. Endogenous DNA proved to be quite stable to even extreme sodium hypochlorite treatments (6% for 21h), suggesting that DNA adsorbs to hydroxyapatite in the bone and that this process facilitates the preservation of DNA in ancient skeletal remains.
机译:从考古或法医骨骼遗骸中提取DNA可以为分析提供非常有力的数据,但受到一系列独特的方法学问题的困扰。在此类分析中要克服的最重要的方法论问题之一是骨骼和牙齿表面存在现代污染,除非在开始DNA提取之前将其清除,否则可能导致假阳性和错误结果。古代DNA(aDNA)研究人员和法医科学家已采用多种技术来最大限度地减少此类污染。一种这样的技术是使用漂白剂(次氯酸钠-NaOCl)“破坏”污染的DNA。但是,尚未就使用的次氯酸钠的最佳浓度以及骨骼或牙齿应接触的时间达成共识。本研究通过将污染引入远古骨骼(从大约500 BP起)并确定几种次氯酸钠处理中哪一种能最好地消除表面污染来系统地解决这一问题。要消除骨骼表面的污染,需要将其浸入至少3.0%(w / v)的次氯酸钠(约等于商业漂白剂和水的等份)中至少15分钟。内源性DNA即使在极端的次氯酸钠处理中(6%,持续21h)也被证明是非常稳定的,这表明DNA可以吸附到骨骼中的羟基磷灰石上,并且该过程有助于DNA在古代骨骼遗骸中的保存。

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