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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。
Abstract：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.
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基本信息:
中图分类号:R446.8;R450
引用信息:
[1]杜靖,古丽孜亚·卡克巴依,海米提·阿布都力木.关于甲醛固定石蜡包埋组织提取DNA质量的探讨[J].新疆医科大学学报,2008(09):1146-1147.
基金信息:
国家自然科学基金资助课题(30460046)
2008-09-30
2008-09-30