法医学杂志

法医学杂志 ›› 2009, Vol. 0 ›› Issue (2): 109-111+.

• 论文 • 上一篇    下一篇

全血中DNA6种提取方法的比较

畅晶晶;张素华;李莉;   

  1. 复旦大学上海医学院法医学系;司法部司法鉴定科学技术研究所上海市法医学重点实验室;苏州大学医学部法医学系;
  • 发布日期:2009-04-25 出版日期:2009-04-28

Comparing and Evaluating Six Methods of Extracting Human Genomic DNA from Whole Blood

CHANG JING-JING1,2, ZHANG SU-HUA2,3, LI LI2 (1. DEPARTMENT OF FORENSIC MEDICINE, SHANGHAI MEDICAL COLLEGE, FUDAN UNIVERSITY, SHANGHAI 200032, CHI- NA; 2. SHANGHAI KEY LABORATORY OF FORENSIC MEDICINE, INSTITUTE OF FORENSIC SCIENCE, MINISTRY OF JUSTICE, P.   

  • Online:2009-04-25 Published:2009-04-28

被引次数

23

摘要： 目的比较经典有机法、改良有机法、常规Chelex-100法、IQ法、Qiagen法及SP法6种方法在提取DNA纯度和得率上的差异。方法收集10名健康志愿者的静脉全血各5mL,分别采用6种方法提取基因组DNA,通过紫外分光光度仪和荧光定量分析技术检测产物的纯度和浓度,计算得率,并使用统计软件对结果进行分析。结果常规Chelex-100法所得DNA的纯度明显低于其他方法,而另外5种方法所得DNA纯度的差异不具有统计学意义。改良有机法得率最低,IQ法得率最高。统计结果表明试剂盒方法抽提全血DNA的得率明显高于经典有机法、常规Chelex-100法和改良有机法,其差异具有统计学意义。结论与有机法和常规Chelex-100法相比,高质量试剂盒类方法更有利于法医学检材的DNA抽提。

关键词: 法医遗传学, 提取法, 试剂盒, 诊断, DNA定量

Abstract: Objective Comparing the differences in purity and yield among six methods of extracting human genomic DNA from whole blood, which included Classic Phenol-chloroform extraction, modified combined technique composed of improved Phenol-chloroform extraction and Chelex-100 extraction, Chelex-100 extraction, IQ, Qiagen and SP. Methods Ten samples of intravenous whole blood(5 mL/sample) were collected and human genomic DNA was extracted with these six methods. The purity and concentration of the DNA products were detected by ultraviolet spectrophotometry and fluorescent quantitation technique, the yield was calculated and tested with statistical software. Results The Chelex-100 extraction was inferior in DNA purity to other methods while the other five methods showed no statistical difference. Modified combined technique was the poorest and IQ was the best in yield among the six methods of extraction. Statistical result showed that the extraction with high quality kits was better than that with classic Phenol-chloroform extraction, Chelex-100 extraction and modified combined technique composed of improved Phenol-chloroform. There was statistical difference between them. Conclusion Comparing to Phenol-chloroform extraction and Chelex-100 extraction, high quality kits are more useful in DNA extraction from forensic materials.

Key words: forensic genetics, extraction, reagent kits, diagnostic, DNA quantitative