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烧骨DNA提取与检测技术的实验研究
引用本文:徐国昌,侯续伟,任甫,刘海东. 烧骨DNA提取与检测技术的实验研究[J]. 中国临床解剖学杂志, 2012, 30(1): 91-95
作者姓名:徐国昌  侯续伟  任甫  刘海东
作者单位:1.南阳理工学院生物人类学研究所, 河南 南阳 473004; 2.辽宁医学院人类学研究所, 辽宁 锦州 121000;
3.潍坊市公安局, 山东 潍坊 261041
基金项目:辽宁省百千万人才工程培养经费资助项目(2010921042)
摘    要:目的 探索烧骨DNA提取与检测方法。 方法 常压下以200℃分别对30例成人股骨焚烧1h后,生物冷冻研磨,EDTA脱钙,改良酚-氯仿提取DNA,光度计检测DNA浓度和纯度;5色荧光标记,PCR复合扩增,基因分析仪毛细管电泳,软件收集数据和分析。 结果 改良酚-氯仿法提取烧骨DNA浓度为(28.5±1.7 )ng/μl,16个基因位点基本齐全,相对荧光单位(RFU)>500;传统酚-氯仿法提取烧骨DNA浓度为(4.7±0.8 )ng/μl,基因位点检出不全,RFU<500。 结论 改良酚-氯仿法、PCR复合扩增、基因分析能对烧骨DNA进行有效提取和检测,但操作时要严格规程。

关 键 词:DNA检验  酚-氯仿法  复合扩增  烧骨  
收稿时间:2011-07-05

Experimental study of DNA extraction and detection in burned bone
XU Guo-chang , HOU Xu-wei , REN Fu , LIU Hai-dong. Experimental study of DNA extraction and detection in burned bone[J]. Chinese Journal of Clinical Anatomy, 2012, 30(1): 91-95
Authors:XU Guo-chang    HOU Xu-wei    REN Fu    LIU Hai-dong
Affiliation:1.Institute of Anthropology, Nanyang Institute of Technology, Nanyang 473004, China; 2. Institute of Anthropology, Liaoning Medical University, Jinzhou 121000, China; 3. Weifang Public Security Bureau, 261041.China
Abstract:Objective To explore how to extract and detect DNA from burned bone.Methods 30 adult femoral bone were incinerated at 200℃ 1 hour in normal pressure,and then freezed and grinded to be bone meal in the liquid nitrogen before decalcification by ethylenediamine tetraacetic acid(EDTA).The deoxyribonucleic acid(DNA) was extracted by improved phenol-chloroform,and the luminometer was used to detect the concentration and purity of the purified DNA.Subsequently DNA was five-fluorescent labeled,amplified by PCR,followed by capillary electrophoresis and the data analysis.Results DNA concentration was(28.5±1.7) ng/μl under the improved phenol-chloroform method,and the 16 genetic locus were detected,with the relative fluorescence units(RFU) of more than 500.The DNA concentration was about(4.7±0.8) ng /μl,and some gene cound not be detected under the traditional phenol-chloroform extraction.Conclusions The DNA extraction and detection can be performed in burned bone by the improved phenol-chloroform method,multiplex polymerase chain reaction,and gene locus analysis.
Keywords:DNA detection  Phenol-chloroform  Multiplex polymerase chain reaction  Burned bone
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