Szoke M, Sasvari-Szekely M, Guttman A
Genetic BioSystems, San Diego, CA 92121, USA.
J Chromatogr A. 1999 Jan 15;830(2):465-71. doi: 10.1016/s0021-9673(98)00878-4.
A novel, rapid and efficient separation method is described for the analysis of double stranded (ds) DNA fragments in the form of horizontal ultra-thin-layer agarose gel electrophoresis. This separation technique combines the multilane, high-throughput separation format of agarose slab gel electrophoresis with the excellent performance of capillary electrophoresis. The electrophoretic separation of the fluorophore (Cy5)-labeled dsDNA molecules were imaged in real time by a scanning laser-induced fluorescence/avalanche photodiode detection system. Effects of the gel concentration (Ferguson plot) and separation temperature (Arrhenius plot) on the migration characteristics of the DNA fragments are discussed. An important genotyping application is also shown by characterizing the polymorphic region (2 X or 4 X 48 base pair repeats) of the dopamine D4 receptor gene (D4DR, exon III region) for ten individuals, using PCR technology with Cy5-labeled primers and ultra-thin-layer agarose gel electrophoresis.
本文描述了一种用于分析双链（ds）DNA片段的新型、快速且高效的分离方法，该方法采用水平超薄层琼脂糖凝胶电泳形式。这种分离技术将琼脂糖平板凝胶电泳的多通道、高通量分离形式与毛细管电泳的卓越性能相结合。通过扫描激光诱导荧光/雪崩光电二极管检测系统对荧光团（Cy5）标记的dsDNA分子的电泳分离进行实时成像。讨论了凝胶浓度（弗格森图）和分离温度（阿累尼乌斯图）对DNA片段迁移特性的影响。还展示了一个重要的基因分型应用，即使用带有Cy5标记引物的PCR技术和超薄层琼脂糖凝胶电泳，对十个人的多巴胺D4受体基因（D4DR，外显子III区域）的多态性区域（2X或4X 48碱基对重复）进行特征分析。
