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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 compare the effects of three DNA extraction methods on formalin-fixed paraffin-embedded autopsy tissues on the quality of DNA, and to find a method for DNA extraction from paraffin-embedded tissues that is easy to operate, less contaminated and less costly. Methods Twenty paraffin-embedded myocardial tissue samples were obtained. The DNA was extracted by xylene dewaxing-phenol-chloroform method, modified TES water-bath dechlorination-phenol-chloroform method, and the kit was used for electrophoresis analysis and UV spectrophotometer A260 / A280 value and PCR amplification. Results The modified TES water bath dewaxing - phenol - chloroform extraction DNA method, xylene dewaxing - phenol chloroform method were compared with the A260 / A280 DNA extracted by the kit method, were significant (P <0.01). There was no significant difference (P> 0.05) between A260 / A280 values of DNA extracted from dewaxing-phenol-chloroform method and modified TES water bath-phenol-chloroform method. The DNA obtained from the dewaxing-phenol-chloroform method in the TES water bath was used as a template, and the brightness of the target band amplified was comparable to that of the positive control. Conclusion The results show that the modified TES water bath dewaxing - phenol chloroform method is simple and effective reagents used inexpensive, is an economical and practical method of DNA extraction of paraffin-embedded tissue.