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通过高压灭菌和紫外线照射消除污染的扩增短串联重复序列产物。

Elimination of contaminating amplified short tandem repeat products by autoclaving and ultraviolet irradiation.

作者信息

Ohta Jun, Tanaka Atsunori

机构信息

Forensic Biology Unit, Scientific Crime Laboratory, Kanagawa Prefectural Police, Japan.

出版信息

Med Sci Law. 2018 Jan;58(1):25-31. doi: 10.1177/0025802417747166. Epub 2017 Dec 28.

Abstract

DNA contamination can result in false interpretation of short tandem repeat (STR) DNA typing. Proper decontamination is particularly required in forensic DNA laboratories where probative value of the evidence may be affected. The aim of this study was to establish an effective DNA decontamination procedure for amplified STR products focusing on laboratory-related contamination. We verified the effectiveness of thermally and temporally extended autoclaving and ultraviolet irradiation for the elimination of contaminating amplified STR products. STR amplification products were prepared using a control genomic DNA template and generated using the AmpFℓSTR Identifiler Plus and Yfiler polymerase chain reaction amplification kits. In this study, the contaminants were dried before decontamination treatment, which resembles actual contamination situations. One microlitre of amplified STR products was eliminated by a combination of autoclaving (128°C, 420 min) and UV irradiation (60 J/cm). Our results reveal that the combination treatment represents an effective DNA decontamination procedure and a practicable method in standard level laboratories. Finally, we propose a comprehensive approach for forensic DNA laboratories to implement to minimise contamination issues and guarantee provision of authentic results.

摘要

DNA污染会导致对短串联重复序列(STR)DNA分型的错误解读。在可能影响证据证明价值的法医DNA实验室中,尤其需要进行适当的去污处理。本研究的目的是建立一种针对扩增后的STR产物的有效DNA去污程序,重点关注与实验室相关的污染。我们验证了热延长和时间延长高压灭菌以及紫外线照射对消除污染的扩增STR产物的有效性。使用对照基因组DNA模板制备STR扩增产物,并使用AmpFℓSTR Identifiler Plus和Yfiler聚合酶链反应扩增试剂盒进行扩增。在本研究中,污染物在去污处理前进行干燥处理,这类似于实际污染情况。通过高压灭菌(128°C,420分钟)和紫外线照射(60 J/cm)相结合的方式消除了1微升扩增的STR产物。我们的结果表明,联合处理是一种有效的DNA去污程序,也是标准水平实验室中一种可行的方法。最后,我们为法医DNA实验室提出了一种全面的方法,以实施该方法来尽量减少污染问题并保证提供可靠的结果。

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