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化学学报
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化学学报 ›› 2004, Vol. 62 ›› Issue (1): 66-70. 上一篇    下一篇

研究论文

毛细管电泳紫外检测条件下单碱基分离DNA片段

王前, 许旭   

  1. 中国科学院上海有机化学研究所分析测试中心, 上海, 200032
  • 投稿日期:2003-05-26 修回日期:2003-09-05 发布日期:2014-01-26
  • 通讯作者: 许旭 E-mail::E-mail:xuxu@mail.sioc.ac.cn.
  • 基金资助:
    中国科学院创新工程重大项目基金(No.KSCX1-06)资助.

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

WANG Qian, XU Xu   

  1. Analytical and Testing Center, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032
  • Received:2003-05-26 Revised:2003-09-05 Published:2014-01-26

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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pBR322/Hae Ⅲ样品进行分离,其中123/124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123/124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.

关键词: 毛细管电泳, 无胶筛分, 合成水溶性聚合物, DNA分离, 紫外检测器, 单碱基分离

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/Hae Ⅲ 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 (Rs>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.

Key words: capillary electrophoresis, non-gel sieving, synthetic polymer, DNA separation, UV detector, single-base-pair separation