Song L, Fang D, Kobos R K, Pace S J, Chu B
Department of Chemistry, State University of New York at Stony Brook, NY 11794-3400, USA.
Electrophoresis. 1999 Oct;20(14):2847-55. doi: 10.1002/(SICI)1522-2683(19991001)20:14<2847::AID-ELPS2847>3.0.CO;2-W.
The separation of double-stranded DNA (dsDNA) fragments in polymethylmethacrylate (PMMA) capillary electrophoresis (CE) chips by using E99P69E99 as a separation medium has been demonstrated. The PMMA CE chips were simply manufactured by micromachining and adhesive tape sealing. To make the separation channel compatible with the separation medium, a dynamic nonionic surfactant coating procedure was developed, which made the plastic separation channel sufficiently hydrophilic to allow the separation medium to fill the channel by capillary action. Subsequent separation of DNA fragments was successful with a separation efficiency of the order of 10(4) theoretical plates over an effective separation distance of 1.5 cm. By using an applied electric field strength of 200 V/cm, the separation of low DNA mass ladder was completed within 5 min. The simple coating procedure, together with the self-assembled viscosity-adjustable separation medium, should be useful to meet some of the essential requirements for developing single-use disposable CE chips. Coating the channels with polymer blends of PMMA and the separation medium also showed promise.
已证明使用E99P69E99作为分离介质在聚甲基丙烯酸甲酯(PMMA)毛细管电泳(CE)芯片中分离双链DNA(dsDNA)片段。PMMA CE芯片通过微加工和胶带密封简单制造。为使分离通道与分离介质兼容,开发了一种动态非离子表面活性剂涂层程序,该程序使塑料分离通道具有足够的亲水性,以允许分离介质通过毛细管作用填充通道。随后成功分离了DNA片段,在1.5 cm的有效分离距离上分离效率约为10⁴理论塔板数。通过使用200 V/cm的施加电场强度,在5分钟内完成了低分子量DNA梯度的分离。这种简单的涂层程序,连同自组装的粘度可调分离介质,应该有助于满足开发一次性使用CE芯片的一些基本要求。用PMMA和分离介质的聚合物共混物涂覆通道也显示出前景。