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目的对不同方法提取甲醛固定组织中DNA的效果进行比较,寻找一种操作简便、经济实用、质量较高的DNA提取方法。方法取甲醛固定的心肌组织14份,分别以改良酚-氯仿法,改良Trizol法,试剂盒法提取DNA,进行紫外分光光度计测定OD260/OD280值后,经PCR扩增,琼脂糖凝胶电泳分析确定提取的DNA质量。结果改良酚-氯仿法,改良Trizol法,试剂盒法OD260/OD280比值分别为1.841 5±0.380 4、1.370 5±0.336 7、0.831 6±0.175 0。两两比较均有显著性差异(P<0.05)。3种不同方法提取DNA含量分别为0.943 8±0.530 1、0.707 5±0.423 6、0.342 8±0.182 5。PCR扩增后琼脂糖凝胶电泳显示以改良酚-氯仿法所提DNA的谱带清晰度好于其它两种方法。结论改良酚-氯仿法简便有效,所用试剂价格低廉,是一种经济实用的甲醛固定组织DNA提取方法。
OBJECTIVE To compare the effect of extracting DNA from formalin-fixed tissues by different methods and to find a DNA extraction method with simple, economical, practical and high quality. Methods Fourteen formaldehyde-fixed myocardial tissues were obtained. The DNA was extracted by modified phenol-chloroform method, modified Trizol method and kit method. The OD260 / OD280 values were determined by ultraviolet spectrophotometer. After PCR amplification, agarose gel electrophoresis Analysis determines the quality of extracted DNA. Results The modified phenol - chloroform method, modified Trizol method, kit OD260 / OD280 ratio were 1.841 5 ± 0.380 4,1.370 5 ± 0.336 7,0.831 6 ± 0.175 0. There was a significant difference between any two groups (P <0.05). The DNA contents extracted by three different methods were 0.943 8 ± 0.530, 0.707 5 ± 0.423 6 and 0.3442 8 ± 0.182, respectively. The agarose gel electrophoresis after PCR amplification showed that the band definition of DNA extracted by modified phenol-chloroform method was better than the other two methods. Conclusion The modified phenol-chloroform method is simple and effective, and the reagent used is cheap. It is an economical and practical method for DNA extraction from fixed-form formaldehyde.