关于甲醛固定石蜡包埋组织提取DNA质量的探讨

Further study on the quality of DNA extracted from formalin fixed and paraffin embedded tissues

目的:探讨甲醛固定石蜡包埋组织(FFPET)提取DNA后PCR扩增产物检测率的提高。方法:应用从FFPET提取的DNA,PCR反应扩增β-Globin(110bp),分析纯化前后DNAPCR产物的检测率。结果:纯化后的DNA模板PCR反应取得高质量目的基因。结论:纯化后能有效去除不能通过蛋白酶K消化和有机萃取法去除的PCR反应抑制因子,利于PCR反应成功。

Objective：Study the effect of DNA purification after extraction from formalin fixed and paraffin embedded tissues（FFPET）on PCR amplification efficiency.Methods：Compared the PCR amplification efficiency of DNA samples extracted from formalin fixed and paraffin embedded tissues and the DNA samples further purified by UNIQ-10 spin column after extraction using specific primers and optimized PCR conditions for β-globulin（110 bp）.Results：Further purification of DNA samples with UNIQ-10 spin column after DNA extraction increased PCR amplification efficiencies for the gene studied in all samples were compared to the DNA samples without further purification step.Conclusion：Further purification of DNA samples with UNIQ-10 spin column after extraction removes the PCR inhibiting factors that could not be removed by proteinase K in the DNA templates from formalin fixed and paraffin embedded tissues,and increases the sensitivity of PCR amplification.