甲醛固定石蜡包埋组织中DNA的三种提取方法比较

Comparison of three methods of DNA extraction fromformaldehyde-fixed, paraffin-embedded tissues

为了寻找最佳的自普通 10 %甲醛固定石蜡包埋组织中提取DNA的方法 ,取我院 2 0 0 0年手术切除的 10 %甲醛固定石蜡包埋的食管癌标本 10例 ,分别以蛋白酶K消化酚氯仿抽提法 ,蛋白酶K消化氯化钠盐析法和不同pH值下加热提取法提取其DNA ,然后进行电泳和PCR扩增分析。结果显示 ,蛋白酶K消化酚氯仿抽提法 ,蛋白酶K消化氯化钠盐析法所得DNA产量和质量均高于不同pH值下加热提取法。从而认为蛋白酶K消化氯化钠盐析法简便快捷 ,不用接触有毒的化学药品 ,是理想的DNA提纯方法。

In order to find an ideal method to extract DNA from unbuffered formalin-fixed, paraffin-embedded tissues, ten blocks of carcinoma of the esophagus resected in our hospital were selected in 2000. The DNAs of these samples were obtained by three methods: extracting by phenol:chloroform after proteinase K digestion, salting-out by NaCl after proteinase K digestion, extracting by phenol:chloroform after heating under the influence of pH. Then, the DNAs were analyzed by electrophoresis and PCR amplification. It showed that the qualities and quantities of DNAs obtained by phenol:chloroform extraction and NaCl salting-out were better than that of heating under different pH. So, we conclude that NaCl salting-out after proteinase K digestion is an ideal method for DNA extraction because of its simplicity and non-toxicity.