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目的探讨通过石蜡包埋组织标本构建肝细胞癌(hepatocellular carcinoma,HCC)肿瘤组织基因组DNA文库的方法。方法收集符合要求的石蜡包埋HCC肿瘤组织标本,运用组织裂解改良法提取基因组DNA,构建HCC肿瘤组织基因组DNA文库,通过SmartSpecTM核酸蛋白测量法、TaqMan-MGB-PCR及琼脂糖凝胶电泳法等分析评估所得文库质量。结果本研究共收集到2 108例合格石蜡包埋HCC肿瘤组织标本,构建了HCC组织标本基因组DNA文库,经质量监控分析所得DNA的浓度大于或接近0.1μg/μl,对应A260/A280比值介于1.6~2.0,TaqMan-MGB-PCR实时定量曲线光滑、波峰单一,琼脂糖凝胶电泳条带单一、清晰、无拖尾现象,所构建的文库DNA质量较好,与对照DNA质量相比差异无显著性(P>0.05)。结论通过石蜡包埋组织标本可构建用于HCC研究的肿瘤组织基因组DNA文库。
Objective To investigate the method of constructing genomic DNA library of hepatocellular carcinoma (HCC) tumor tissue by paraffin-embedded tissue samples. Methods The samples of paraffin-embedded HCC tumor tissues were collected. The genomic DNA was extracted by tissue lysis method. The genomic DNA library of HCC tumor tissues was constructed. By using SmartSpec ™ nucleic acid protein assay, TaqMan-MGB-PCR and agarose gel electrophoresis Analyze and evaluate the quality of the resulting library. Results A total of 2 108 specimens of paraffin-embedded HCC tumor tissues were collected and the genomic DNA library of HCC tissue samples were constructed. The DNA concentration of the HCC tissue samples was greater than or close to 0.1 μg / μl, corresponding to A260 / A280 ratio 1.6 ~ 2.0, TaqMan-MGB-PCR real-time quantitative curve is smooth, single peak, agarose gel electrophoresis stripe with a single, clear, no tailing, the construction of the library DNA quality is better, compared with the control DNA quality difference Significance (P> 0.05). Conclusion Tumor tissue genomic DNA libraries for HCC studies can be constructed by paraffin-embedded tissue specimens.