法医学杂志

法医学杂志 ›› 2009, Vol. 0 ›› Issue (6): 421-424.

• 论文 • 上一篇    下一篇

运用酚-氯仿法结合磁珠法提取蝇蛆体内人类DNA

王翔;蔡继峰;钟鸣;李剑波;张金国;兰玲梅;   

  1. 中南大学基础医学院法医学系;中南大学湘雅二医院;长沙市公安局;
  • 发布日期:2009-12-25 出版日期:2009-12-28

Extract Human DNA from Maggot Crop Contents by Phenol-chloroform Method Coupled with Paramagnetic Particle Method

WANG XIANG1, CAI JI-FENG1,2, ZHONG MING1, LI JIAN-BO3, ZHANG JIN-GUO3, LAN LING-MEI1 (1. DEPARTMENT OF FORENSIC MEDICINE, COLLEGE OF PRECLINICAL MEDICINE, CENTRAL SOUTH UNIVERSITY, CHANGSHA 410013, CHINA; 2. THE SECOND XIANGYA HOSPITAL OF CENTRAL SOUTH UN   

  • Online:2009-12-25 Published:2009-12-28

被引次数

4

摘要： 目的建立运用酚-氯仿法结合磁珠法从蝇蛆嗉囊内提取人类DNA的方法,从而提高STR分型检验的灵敏度。方法采用酚-氯仿法对蝇蛆嗉囊内容物中的人类DNA进行提取,提取产物经磁珠法纯化浓缩后用QuantifilerTM人类DNA定量试剂盒在7500型实时荧光定量PCR仪上进行PCR定量,再用AmpF■STR IndentifilerTM试剂盒在3130XL-Avant遗传分析仪上对这些DNA样本进行STR分型。结果本研究建立的方法可增加模板DNA浓度约为单独使用酚-氯仿法的2倍。用此方法提取到的DNA浓度[(0.218±0.041)ng/μL]可获得全部16个STR分型结果。结论酚-氯仿法结合磁珠法可以有效地提高提取到的人类DNA样本STR分型检验的灵敏度,对于从事法医昆虫学方面研究的工作者有较好的实用价值。

关键词: 法医遗传学, 昆虫学, 提取法, 酚-氯仿法, 磁珠法, 蝇科

Abstract: Objective To establish an effective phenol-chloroform method coupled with paramagnetic particle method for human DNA extraction from maggot crop contents in STR genotyping. Methods Human DNA was extracted from the maggot crop contents using phenol-chloroform method and purified by para- magnetic particle method. DNA was quantified by PCR with QuantifilerTM Human DNA Quantification Kit using 7500 real-time fluorescence quantitative PCR instrument. PCR products were genotyped by AmpF■STR IdentifilerTM PCR Amplification Kit using 3130XL-Avant genetic analyzer. Results The template DNA yield by the method described were increased at least 2 times than the phenol-chloroform extraction method alone. All of the full 16 STR profiles could be obtained with the samples extracted by this method when the DNA yield reached (0.218±0.041) ng/μL. Conclusion Phenol-chloroform method coupled with paramagnetic particle method can effectively increase the sensitivity of STR analysis of human DNA recovered from maggot crop contents and is a valuable tool for forensic entomology.

Key words: forensic genetics, entomology, extraction method, phenol-chloroform method, paramagnetic particle method, muscidae