Paulus A, Hüsken D
Corporate Analytical Research, Ciba-Geigy Ltd., Basal, Switzerland.
Electrophoresis. 1993 Jan-Feb;14(1-2):27-35. doi: 10.1002/elps.1150140106.
DNA digest analysis with polymer-filled capillaries in capillary electrophoresis is described. The samples analyzed consisted of commercially available standards including a 100 base pair ladder with repeating units up to 2000 base pairs. Three DNA digests covered the most common small fragment ranges: up to 600 base pairs, up to 1500 base pairs and from 100 to 2500 base pairs. All samples were separated by traditional slab gel electrophoresis at various agarose concentrations and by automated capillary electrophoresis. The capillary electrophoretic separations were achieved with noncross-linked polyacrylamide from 6% monomer solutions. The acrylamide was polymerized inside the capillary, which was coated with a methacryloxysilane to insure binding of the polyacrylamide to the capillary wall. With 6% columns, excellent separations were observed up to 600 base pairs with base line resolution for fragments differing in less than 10 base pairs. Resolution power for fragments between 600 and 2200 decreased to about 300 base pairs. Compared to slab gels, capillary electrophoresis achieved better resolution in the low fragment range, whereas with the reported column composition, slab gels were superior above 600 base pairs. Fast access to the analysis of a specific sample, automation, and a larger dynamic range for sample load are further benefits of a DNA digest analysis by capillary electrophoresis.
描述了在毛细管电泳中使用填充聚合物的毛细管进行DNA消化分析。所分析的样品包括市售标准品，其中有一个100碱基对的梯度标准，重复单元可达2000碱基对。三种DNA消化产物覆盖了最常见的小片段范围：高达600碱基对、高达1500碱基对以及100至2500碱基对。所有样品都通过不同琼脂糖浓度的传统平板凝胶电泳和自动毛细管电泳进行分离。毛细管电泳分离是用6%单体溶液的非交联聚丙烯酰胺实现的。丙烯酰胺在毛细管内聚合，毛细管涂有甲基丙烯酰氧基硅烷以确保聚丙烯酰胺与毛细管壁结合。使用6%的柱，在高达600碱基对时观察到了出色的分离效果，对于相差不到10个碱基对的片段具有基线分辨率。600至2200碱基对之间片段的分辨率降至约300碱基对。与平板凝胶相比，毛细管电泳在低片段范围内实现了更好的分辨率，而对于所报道的柱组成，平板凝胶在600碱基对以上更具优势。快速进行特定样品分析、自动化以及更大的样品加载动态范围是毛细管电泳进行DNA消化分析的进一步优势。
