High-frequency alternating-crossed-field gel electrophoresis with neutral or slightly charged interpenetrating networks to improve DNA separation.
High-frequency alternating-crossed-field gel electrophoresis with neutral or slightly charged interpenetrating networks to improve DNA separation.
复制标题
采用中性或微带电互穿网络的高频交替交叉场凝胶电泳可改善 DNA 分离。
DOI：
10.1002/elps.1150191815
复制
发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Blanch,HW
中科院分区：
文献类型：
--
作者：
Boyd,BM;Prausnitz,JM;Blanch,HW
为了改善 DNA 分离，这项工作报告了垂直于直流 (DC) 分离场叠加的高频方波交流场对基于聚丙烯酰胺的互穿网络 (IPN) 和琼脂糖网络中 DNA 迁移的影响。与标准聚丙烯酰胺凝胶相比，IPN 可以分离较大的 DNA（5 V/cm 时为 9000 bp 与 5000 bp）。在新型聚丙烯酰胺基 IPN 中，5 Hz 的交流 (AC) 场增加了可分离的最大 DNA 尺寸。随着电场强度的增加，这种效应扩展到更大的 DNA 尺寸，直至并明显超过电源限制的最大电场强度 48 V/cm。正交交流场还增加了移动性。这两个结果相结合，使基于聚丙烯酰胺的新型 IPN 的分离时间缩短了 20 倍。当带负电荷的丙烯酸基团被纳入 IPN 中时，AC 场的使用改变了 DNA 网络相互作用，从而改变了 DNA 迁移率的大小依赖性。在琼脂糖凝胶中，50 Hz 的交流场增加了可分离的尺寸范围；然而，DNA 迁移率并没有增加。当琼脂糖网络中带电基团的数量增加时，交流场中迁移率的大小依赖性没有变化。根据文献中的结果，研究了交流场对 DNA 分离影响的可能机制。
Toward improving DNA separations, this work reports the effects of high‐frequency square‐wave AC fields superimposed perpendicular to the direct current (DC) separation field on DNA migration in both polyacrylamide‐based interpenetrating networks (IPNs) and in agarose networks. Compared to standard polyacrylamide gels, IPNs allow the separation of larger DNA (9000 bpvs.5000 bp at 5 V/cm). In novel polyacrylamide‐based IPNs, an alternating current (AC) field of 5 Hz increased the maximum DNA size separable. This effect was extended to larger DNA sizes with increasing electric‐field strength up to and apparently beyond the power supply‐limited maximum electric‐field strength of 48 V/cm. The orthogonal AC field also increased mobility. These two results combine to yield a reduction in separation time of up to a factor of 20 in novel polyacrylamide‐based IPNs. When negatively charged acrylic‐acid groups were incorporated into the IPNs, the use of the AC field changed the DNA‐network interaction, which altered the size dependence of DNA mobility. In agarose gels, an AC field of 50 Hz increased the size range separable; however, there was no increase in DNA mobility. There was no change in size dependence of mobility in an AC field when the number of charged groups in the agarose network was increased. Based on results in the literature, possible mechanisms were examined for the effects of the AC field on DNA separation.