烧骨DNA提取与检测技术的实验研究

中国临床解剖学杂志 ›› 2012, Vol. 30 ›› Issue (1): 91-95.

烧骨DNA提取与检测技术的实验研究

徐国昌¹，　侯续伟²,　任　甫²，　刘海东³

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

收稿日期:2011-07-05 出版日期:2012-01-25 发布日期:2012-02-03
通讯作者: 任　甫，教授，硕士生导师，E-mail:anatomy@yeah.net E-mail:xuguoch@163.com
作者简介:徐国昌(1971-)，男，河南南阳人，硕士，副教授，从事人类学及应用解剖学研究
基金资助:
辽宁省百千万人才工程培养经费资助项目(2010921042)

Experimental study of DNA extraction and detection in burned bone

XU Guo-chang¹，HOU Xu-wei², REN Fu²，LIU Hai-dong³

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

Received:2011-07-05 Online:2012-01-25 Published:2012-02-03

摘要/Abstract

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

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.

Key words: DNA detection, Phenol-chloroform, Multiplex polymerase chain reaction, Burned bone

中图分类号:

R89
DF795.6

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.

XU Guo-Chang, HOU Xu-Wei, LIN 　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.