摘要
目的比较甲醛固定石蜡包埋尸检组织中三种提取DNA的方法对DNA质量的影响,寻找一种操作简便、污染较少、经济实用的石蜡包埋组织中提取DNA的方法。方法选取经甲醛固定石蜡包埋的心肌组织20例,分别以二甲苯脱蜡-酚氯仿法、改良TES水浴脱蜡-酚氯仿法、试剂盒法提取DNA,进行电泳分析、紫外分光光度计测定A260/A280值及PCR扩增。结果改良TES水浴脱蜡-酚氯仿法抽提DNA法、二甲苯脱蜡-酚氯仿法分别与试剂盒法所提取DNA的A260/A280值相比较,均有显著性意义(P<0.01)。二甲苯脱蜡-酚氯仿法、改良TES水浴脱蜡-酚氯仿法所提取DNA的A260/A280值相比较,无显著性意义(P>0.05)。以改良TES水浴脱蜡-酚氯仿法所得DNA为模板,扩增的目的条带亮度与阳性对照相当。结论结果证实改良TES水浴脱蜡-酚氯仿法简便有效,所用试剂价格低廉,是一种经济实用的石蜡包埋组织DNA提取方法。
Objective To investigate the influence of three DNA extraction methods for formalin-fixed and paraffin-embedded autopsy tissue on DNA quality,and look for a simple,less pollution,economic and practical method for DNA extraction from paraffin-embedded autopsy tissue.Methods 20 cases of formalin-fixed and paraffin-embedded myocardial tissue were selected and divided into three groups according to the DNA extraction methods including xylenedewaxing and phenol-chloroform,improved TES dewaxing with water-bath and phenol-chloroform,and DNA extraction kit methods.The quality of obtained DNA was analyzed by electrophoresis and PCR amplification.Results There were significant difference in the A260/A280 ratio（P0.01） between the first two methods and the DNA extraction kit method,and no significant difference was observed between the improved TES dewaxing with water-bath and phenol-chloroform method and the xylenedewaxing and phenol-chloroform method（P0.05）.When the extracts with the improved TES dewaxing with water-bath and phenol-chloroform method were used as template,the target bands were as bright as that of the positive control.Conclusion The results demonstrated that the improved TES dewaxing with water-bath and phenol-chloroform method is an optimal method for DNA extraction from the formaldehyde fixed and paraffin embedded tissues.
出处
《中国法医学杂志》
CSCD
北大核心
2011年第6期465-467,共3页
Chinese Journal of Forensic Medicine
关键词
法医病理学
甲醛
石蜡包埋组织
DNA提取
forensic pathology
formaldehyde
paraffin-embedded tissue
DNA extraction
