Barron A E, Sunada W M, Blanch H W
Department of Chemical Engineering, University of California, Berkeley 94720, USA.
Electrophoresis. 1996 Apr;17(4):744-57. doi: 10.1002/elps.1150170421.
Low-viscosity, aqueous solutions of hydrophilic linear polymers have been shown to be useful for the separation of DNA restriction fragments by capillary electrophoresis (CE). However, the choice of polymer type, size, and concentration remains largely empirical, because the mechanism of high-field electrophoretic DNA separations in polymer solutions is not well understood. To assist in elucidating the mechanism of DNA separation, we experimentally investigated the effects of polymer properties such as stiffness (persistence length), average molecular mass, polydispersity, and hydrophilicity on the separation of DNA ranging from 72 bp to 23 kbp. This was accomplished by comparing the results of DNA separations obtained by counter-migration CE in dilute and semidilute solutions of linear polyacrylamide (PAA), hydroxyethyl-cellulose (HEC), and hydroxypropylcellulose (HPC) polymers of several different average molecular masses.
亲水性线性聚合物的低粘度水溶液已被证明可用于通过毛细管电泳(CE)分离DNA限制片段。然而,聚合物类型、大小和浓度的选择在很大程度上仍然是凭经验的,因为聚合物溶液中高场电泳DNA分离的机制尚未得到很好的理解。为了有助于阐明DNA分离的机制,我们通过实验研究了聚合物特性(如刚性(持久长度)、平均分子量、多分散性和亲水性)对72 bp至23 kbp DNA分离的影响。这是通过比较在几种不同平均分子量的线性聚丙烯酰胺(PAA)、羟乙基纤维素(HEC)和羟丙基纤维素(HPC)聚合物的稀溶液和半稀溶液中通过反向迁移CE获得的DNA分离结果来实现的。