【摘要】 目的:设计并制备65bp-500bp范围的荧光标记DNA分子量内标。方法:p MD18-T Vector连接任意一段割胶回收的DNA片段,克隆后提取重组质粒经双酶切后作为模板,在其上游设计一条固定引物并用荧光标记,在其下游设计一系列引物,经PCR扩增后在ABI 3100遗传分析仪上检测。结果:扩增产物DNA片段大小分别为65bp、105bp、149bp、200bp、241bp、269bp、311bp、345bp、400bp、450bp、500bp,与预期片段大小一致。结论:11个片段经混合调平后,峰型良好,出峰位置均匀,可用于毛细管电泳中DNA片段大小的确定。更多还原

【Abstract】 Objective: To prepare the fluorescently labeled DNA internal lane standard range from 65 bp to 500 bp. Method: Clone and double digestion methods were employed to prepare the template of p MD18-T vector ligated a recovery DNA fragment, a fixed and fluorescently labeled forward primer and common reverse primers were designed, and PCR products were analyzed by PCR amplification and ABI 3100 Genetic Analyzer. Result: The size of PCR products is 65bp、105bp、149bp、200bp、241bp、269bp、311bp、345bp、400bp、450bp、500bp,in line with the expected fragment size. Conclusion: The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.更多还原