Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries.
Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries.
复制标题
使用涂层熔融石英毛细管通过毛细管电泳分离 DNA 限制性片段。
作者：
M. Strege;A. Lagu
研究了几种不同的毛细管电泳技术分离DNA限制性内切片段的能力，最长可达23,000 bp。在未经处理的硅胶毛细管中使用电渗透流的方法最多只能提供1 kbp DNA阶梯中23个片段的部分分辨率。通过在硅胶毛细管内壁涂覆聚丙烯酰胺，并用含有甲基纤维素作为套管介质的缓冲液填充这些毛细管，分离了1 kbp DNA阶梯中的所有片段。此外，该技术还有助于分离lambda DNA-HindIII酶解中非常大的片段。在低分离电位下，使用含有至少0.5%甲基纤维素的缓冲液获得最佳分辨率。该技术的性能，即分辨率和定量，使毛细管电泳成为平板凝胶电泳的有力补充，并可能使其成为琼脂糖凝胶电泳和HPLC的首选替代品，用于确认质粒完整性等应用。
The abilities of several different capillary electrophoresis techniques to separate DNA restriction fragments up to 23,000 bp were investigated. Methods employing electroosmotic flow in an untreated silica capillary were found to provide, at best, only partial resolution of the 23 fragments in a 1-kbp DNA ladder. By coating the inner walls of a silica capillary with poly(acrylamide) and filling these capillaries with buffers containing methylcellulose as a sleving medium, all fragments in the 1-kbp DNA ladder were separated. In addition, this technique facilitated the separation of the very large fragments in a lambda DNA-HindIII digest. Optimum resolution was obtained at low separation potentials using buffers containing at least 0.5% methylcellulose. The performance of this technique, i.e., resolution and quantitation, make capillary electrophoresis a powerful complement to slab gel electrophoresis and may make it a preferred alternative to both agarose gel electrophoresis and HPLC for applications such as the confirmation of plasmid integrity.
影响因子：
7.4
作者：
COBB, KA;DOLNIK, V;NOVOTNY, M
通讯作者：
NOVOTNY, M