SEPARATION MEDIA FOR DNA CAPILLARY/CHIP ELECTROPHORESIS

描述(改编自《调查员摘要》)：的具体目的 更新建议涉及新技术的进一步开发和优化 和更好的分离介质，可以克服许多相关的问题 到今天的聚丙烯酰胺溶液或凝胶：聚合物溶液是 通常太粘而不能填充在毛细管中；聚合在 原位方法不是那么可重复的；聚合物凝胶的保质期是 有限的。关键的方案是利用自组装行为 共聚胶体。 研究人员已经实现了设计聚合物的初步目标 基于EPE型嵌段共聚物(E)自组装行为的网络 和P分别表示聚氧乙烯和聚氧丙烯) 以下性质：(1)在10-15℃时，聚合物溶液是低 1xTBE缓冲液中的粘度液体(小于30厘泊)，便于 填充和取出窄口径毛细管中的分离介质； (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(用一种筛分测试到14个碱基 单链(Ss)DNA片段的碱基分辨率高达1千碱基 具有一个碱基对分辨率的双链DNA片段)；以及(3) 聚合物本身已经成功地充当了一种动态涂层材料。 通过将毛细管电泳法结果与结构研究相关联 使用各种物理技术，如激光散射， 小角X射线散射，小角中子散射，核磁共振， 流变学、粘度学和原子力显微镜的优化 用于DNA毛细管电泳的自组装聚合物网络可以 已实现。除了EPE、PEP、EBE和BEB型三块(带B 由于是聚氧丁烯，并且比P)更疏水，建议使用 结晶和线圈崩塌作为物理现象能够 依赖温度的可逆交联化形成。这个 结晶方法特别吸引人，因为超分子 结构可以更具弹性，并且不处于动态平衡状态 单链聚合物，如在自组装过程中。

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.