IMPROVED SEPARATION MEDIA FOR ELECTROPHORESIS

描述（改编自研究者摘要）： 更新建议涉及进一步发展和优化新的 和更好的分离介质，可以克服许多问题， 到目前的聚丙烯酰胺溶液或凝胶：聚合物溶液， 通常太粘而不能填充在毛细管中; 原位方法不是如此可再现的;聚合物凝胶的保存期 有限公司 关键的方案是利用自组装行为 共聚物胶体。 研究人员已经实现了设计一种聚合物的最初目标 基于EPE型嵌段共聚物（E 和P分别表示聚氧乙烯和聚氧丙烯）， 以下性质：（1）在10-15 ℃下，聚合物溶液是低粘度的， 粘度流体（小于30厘泊）在1xTBE缓冲液中， 在细孔毛细管中填充和去除分离介质; (2)在室温（大于20摄氏度）下，聚合物溶液 变成凝胶状，具有良好的筛分能力（用一种碱测试低至14种碱， 单链（ss）DNA片段和最多1个双链酶的碱基分辨率 对于具有一个碱基对分辨率的双链DNA片段， 聚合物本身已经成功地用作动态涂层材料。 通过将毛细管电泳结果与结构研究相关联 使用各种物理技术，例如激光散射， 小角X射线散射，小角中子散射，NMR， 流变仪，粘度计和原子力显微镜，优化的 用于DNA毛细管电泳的自组装聚合物网络可以 办妥了一批 除了EPE、PEP、EBE和BE B型三嵌段（具有B 是聚氧化丁烯并且比P更疏水，建议使用 结晶和线圈塌陷作为物理现象， 温度依赖性可逆交联形成。 的 结晶方法特别有吸引力，因为超分子 结构可以更有弹性， 如在自组装过程中的单个聚合物链。

DESCRIPTION (Adapted from the Investigator's Abstract): The specific aim of the renewal proposal deals with further development and optimization of new and better separation media which can overcome many of the problems related to the present-day polyacrylamide solutions or gels: polymer solutions are often too viscous to be filled in capillary tubings; the polymerization in situ approach is not so reproducible; shelf-life for polymer gels is limited. The key scheme is to take advantage of the self-assembly behavior of copolymer colloids. The investigators have achieved the initial goals of designing a polymer network based on the self-assembly behavior of a EPE-type block copolymer (E and P denoted polyoxyethylene and polyoxypropylene, respectively) with the following properties: (1) at 10-15 degrees C, the polymer solution is a low viscosity fluid (less than 30 centipoise) in 1xTBE buffer, for ease of filling and removing the separation medium in narrow-bore capillary tubings; (2) at room temperatures (greater than 20 degrees C), the polymer solution becomes gel-like with good sieving ability (tested down to 14 bases with one base resolution for single stranded (ss) DNA fragments and up to 1 kilobase pairs for ds DNA fragments with one base pair resolutions); and (3) the polymer itself has acted successfully as a dynamic coating material. By correlating the capillary electrophoresis results with structural studies using a variety of physical techniques, such as laser light scattering, small angle X-ray scattering, small angle neutron scattering, NMR, rheometry, viscosimetry and atomic force microscopy, an optimization of the self-assembled polymer network for DNA capillary electrophoresis can be achieved. In addition to the EPE, PEP, EBE, and BEB type triblocks (with B being polyoxybutylene and more hydrophobic than P), it is proposed to use crystallization and coil-collapse as the physical phenomena capable of temperature-dependent reversible cross-linking formation. The crystallization approach is especially attractive, since the supramolecular structure can be more resilient and is not in dynamic equilibrium with single polymer chains as in the self-assembly process.