Liu T, Liang D, Song L, Nace V M, Chu B
Department of Chemistry, State University of New York at Stony Brook, 11794-3400, USA.
Electrophoresis. 2001 Feb;22(3):449-58. doi: 10.1002/1522-2683(200102)22:3<449::AID-ELPS449>3.0.CO;2-T.
A mixture of two polyoxybutylene-polyoxyethylene-polyoxybutylene (BEB) triblock copolymers (B6E46B6 and B10E271B10, respectively) was used as a new separation medium for separating double-stranded DNA (dsDNA) fragments by capillary electrophoresis (CE). The two block copolymer mixtures were designed to form mixed flower-like micelles in dilute solution and a homogeneous gel-like open-network with hydrophobic clusters as cross-linking points at higher polymer concentrations. Being a polyoxyalkylene block copolymer gel, the separation medium has some special advantages, including the temperature-dependent sol-gel transition that makes sample injection easy, and the self-coating of the inner capillary wall that makes experimental procedures simple and reproducible. Furthermore, it can shorten the elution time and further improve the separation resolution, especially for small dsDNA fragments, when compared with EPE-type separation media, e.g., F127 (E99P69E99, with P being polyoxypropylene) block copolymer gels formed by the closed packing of spherical micelles. Single base pair resolution can be achieved by using the new separation medium for dsDNA fragments up to over 100 base pairs.
两种聚氧丁烯-聚氧乙烯-聚氧丁烯(BEB)三嵌段共聚物(分别为B6E46B6和B10E271B10)的混合物被用作一种新型分离介质,用于通过毛细管电泳(CE)分离双链DNA(dsDNA)片段。设计这两种嵌段共聚物混合物,使其在稀溶液中形成混合花状胶束,并在较高聚合物浓度下形成以疏水簇为交联点的均匀凝胶状开放网络。作为一种聚氧化烯嵌段共聚物凝胶,该分离介质具有一些特殊优势,包括温度依赖性溶胶-凝胶转变,这使得进样容易,以及毛细管内壁的自涂层,这使得实验过程简单且可重复。此外,与EPE型分离介质(例如由球形胶束紧密堆积形成的F127(E99P69E99,其中P为聚氧化丙烯)嵌段共聚物凝胶)相比,它可以缩短洗脱时间并进一步提高分离分辨率,特别是对于小的dsDNA片段。使用这种新型分离介质分离长达100多个碱基对的dsDNA片段时,可以实现单碱基对分辨率。