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毛细管电泳紫外检测条件下单碱基分离DNA片段

DNA Separation as Single-base-pair Resolution by Capillary Electrophoresis with Ultraviolet Detector

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【作者】 王前许旭

【Author】 WANG, Qian XU, Xu (Analytical and Testing Center, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032)

【机构】 中国科学院上海有机化学研究所分析测试中心中国科学院上海有机化学研究所分析测试中心 上海200032上海200032

【摘要】 合成了一种水溶性聚合物 ,聚N ,N 二甲基丙烯酰胺 (PDMA)和聚乙烯吡咯烷酮体系 (PVP) ,并用于DNA分离 .优选该体系溶液含量为 2 0 %PDMA + 2 0 %PVP ,在紫外检测的条件下 ,对标准DNA酶切片段pBR3 2 2 /HaeIII样品进行分离 ,其中 12 3 / 12 4碱基对的分离度 (Rs)为 0 72 .加入甲酰胺 (φ≥ 0 10 )对DNA样品进行去活处理后 ,该体系亦能对单链DNA片段中的 12 3 / 12 4进行分离 ,分离度 (Rs)为 0 75 .实验还显示进样时存在浓缩效应 .本体系粘度较小 ,筛分能力强 ,具备芯片毛细管DNA分离介质的潜力 .

【Abstract】 The water-soluble mixtures of polymers, poly(N,N-dimethylacrylamide) (PDMA)/polyvinylpyrrolidone (PVP), were synthesized and used in capillary electrophoresis (CE) for DNA separation. With the optimal concentration of PDMA/PVP polymer solution, the doublet of 123/124 bp in pBR322/HaeIII Markers could be separated with UV detector and the resolution value was 0.72. When the denaturant formamide (φ≥0.10) was added in the polymer solution, DNA fragments of 123/124 could be also separated well with the resolution as 0.75, which quite met the requirement of DNA sequencing (R s>0.59). In addition, the lower viscosity of the polymer solution made the practice easier. The sample stacking effect was validated in the experiments. The medium showed single-base-pair DNA separation by capillary electrophoresis even with ultraviolet detector and uncoated capillary column.

【基金】 中国科学院创新工程重大项目基金 (No.KSCX1 0 6)资助
  • 【文献出处】 化学学报 ,Acta Chimica Sinica , 编辑部邮箱 ,2004年01期
  • 【分类号】O658.9
  • 【被引频次】8
  • 【下载频次】294
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