石蜡包埋乳腺癌组织DNA提取方法

Paraffin-embedded Breast Cancer Tissue DNA Extraction Method

目的探讨一种乳腺癌患者石蜡包埋组织乳腺癌组织DNA提取方法并进行验证。方法选择无自溶、退变或组织结构不清、大片出血的70例10ml／dl甲醛固定石蜡包埋乳腺癌组织，在避免交叉污染的情况下，经过TES溶液水浴脱蜡后，先后用PBS缓冲液（pH7．2）及无水、95ml／dl，75ml／dl浓度梯度的乙醇60℃温浴4h；加入含蛋白酶K的裂解液56℃消化72h后，酚氯仿法抽提DNA。通过聚合酶链反应（PCR）、琼脂糖凝胶电泳及紫外分光光度仪测定A260nm／A280nm值，分析所提取DNA的质量。结果62例（88．57％）石蜡组织标本DNA获得成功提取，所得DNA样本A260nm／A280nm比值为1．78±0．16，采用此条件处理的样本可获得足够数量的完整DNA，抽提率高，可满足后续临床研究需要。结论改进的石蜡包埋组织提取DNA方法简单、经济，是较理想的非试剂盒法石蜡包埋组织DNA提取方法。

Objective Explore DNA extraction methods of the paraffin-embedded tissue in breast cancer patients and verified it. Methods 70 cases of large areas of bleeding in 10 ml/dl formalin fixed paraffin-embedded breast cancer tissue with avoid autolysis, degeneration,or organizational structure was unclear, and prevent cross-contamination. After TES solution water bathed dewaxing,successively with PBS buffer （pH7.2） and 100 ml/dl, 95 ml/dl, 75 ml/dl ethanol concentration gradient of 60℃ incubation of 4 hours, Phenol-chloroform extraction DNA. By polymerase chain reaction （PCR）, agarose gel electropho- resis and ultraviolet spectrophotometry to measured the A260nm/ A280nm value analysis of the quality of the extracted DNA. Re- suits 62 cases （88.57%） paraffin tissue specimen DNA successfully extracted obtained DNA samples from the A260nm/ A280nm ratio （1.78±0. 16）, This condition of the sample availability of a sufficient number of complete DNA extraction rate, to meet the follow-up clinical research needs. Conclusions The method improved extracting DNA from paraffin-embedded simple,economic and it is ideal means to extract DNA from paraffin-embedded tissues.