Cheng J, Kasuga T, Mitchelson K R, Lightly E R, Watson N D, Martin W J, Atkinson D
Department of Molecular and Cell Biology, University of Aberdeen, UK.
J Chromatogr A. 1994 Aug 12;677(1):169-77. doi: 10.1016/0021-9673(94)80556-3.
Heteroduplex DNA polymorphism analysis (HPA) makes use of conformational polymorphisms to alter electrophoretic mobility of fragments and can be used to detect non-restrictable loci. We have developed a novel application of entangled solution capillary electrophoresis (ESCE) to separate heteroduplex and homoduplex DNA molecules. The addition of ethidium bromide and glycerol to the free solution sieving buffer resulted in the improved peak resolution and good reproducibility. Reannealed polymerase chain reaction products could be used directly for mutation screening and with fully automated ESCE the entire HPA may be completed in less than 30 min including sample handling. This technology could provide a rapid and highly efficient way for screening rare mutations among large numbers of individuals.
异源双链DNA多态性分析（HPA）利用构象多态性来改变片段的电泳迁移率，可用于检测不可限制的基因座。我们开发了一种新型的缠结溶液毛细管电泳（ESCE）应用，用于分离异源双链和同源双链DNA分子。向自由溶液筛分缓冲液中添加溴化乙锭和甘油可提高峰分辨率并具有良好的重现性。重退火的聚合酶链反应产物可直接用于突变筛查，并且通过全自动ESCE，整个HPA包括样品处理在内可在不到30分钟内完成。这项技术可为在大量个体中筛查罕见突变提供一种快速且高效的方法。
