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| 生物检材DNA碱性裂解提取法的优化与改良 | |
| 引用本文： | 李爱强,赵兴春,张艳霞,赵丽,王华,王项华,宋金平.生物检材DNA碱性裂解提取法的优化与改良[J].中国法医学杂志,2011,26(4):301-304. |
| 作者姓名： | 李爱强 赵兴春 张艳霞 赵丽 王华 王项华 宋金平 |
| 作者单位： | 1. 河北省公安厅,河北石家庄,050051 2. 公安部物证鉴定中心,北京,100038 3. 宁夏自治区公安厅,宁夏银川,750000 |
| 摘 要： | 目的通过对DNA碱性裂解提取法进行优化和改进,建立一种操作更简便、检验快速、结果更可靠的生物检材DNA提取方法。方法以抗凝血作为检验样本,通过改变提取试剂的浓度、pH值、保存时间及孵化样本的温度、时间等条件,观测对检验结果峰值的影响,以确定提取试剂最佳条件,DNA提取物最佳保存条件。通过用改良后的碱性裂解法及Chelex100法同时提取不同量的抗凝血样本、各类法医生物检材,用不同厂家的DNA试剂盒扩增,对碱性裂解法的灵敏度、适应性和适用性进行评估。结果改良后的碱性裂解法只需将生物检材用NaOH（0.25mol/L）99℃孵化8min,振荡后加入TrisHCl（0.05mol/L,pH=6.5）中和液,离心后直接扩增检测。DNA提取物于-20℃冰箱可长期保存。对于毛发及精斑类检材优于Chelex100法。DNA提取物适用于现有各种DNA试剂盒扩增检测,其灵敏度与Chelex100法相似。结论改良后的碱性裂解法,操作简便、检验快速、结果可靠,可适合于法庭科学的检案与建库。 |
| 关 键 词： | 法医物证学 碱性裂解法 DNA提取 Chelex100 |
Optimization and Improvement of the Alkaline Lysis Method for the extraction of DNA from Forensic Biological Samples |
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| LI Aiqiang,ZHAO Xingchun,ZHANG Yanxia,ZHAO Li,WANG Hua,WANG Xianghua,SONG Jinping.Optimization and Improvement of the Alkaline Lysis Method for the extraction of DNA from Forensic Biological Samples[J].Chinese Journal of Forensic Medicine,2011,26(4):301-304. | |
| Authors: | LI Aiqiang ZHAO Xingchun ZHANG Yanxia ZHAO Li WANG Hua WANG Xianghua SONG Jinping |
| Institution: | LI Aiqiang1,ZHAO Xingchun2,ZHANG Yanxia1,ZHAO Li1,WANG Hua3,WANG Xianghua1,SONG Jinping1/1.Public Security Bureau of Hebei Province,Shijiazhuang 050051,China,2.Institute of Forensic Science,Ministry of Pubic Security,Beijing 100038,3.Public Security Bureau of Ningxia Province,Yinchuan 750000 |
| Abstract: | Objective To establish a simple,fast and reliable method for extracting DNA from forensically relevant specimens through optimization and improvement of the alkaline lysis protocol.Methods With the anticoagulated blood as test samples,the optimal conditions of the extraction reagents were determined according to the influence on the results of DNA typing of different conditions,including different reagent concentration,pH value and storage time,and different incubation temperature and time.Meanwhile,the opt... |
| Keywords: | forensic biological evidence alkaline lysis method extraction DNA Chelex100 |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |