IMPROVED SEPARATION MEDIA FOR ELECTROPHORESIS

改进的电泳分离介质

• 批准号: 2519138
• 负责人: Benjamin Chu
• 金额: $ 13.18万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2519138
• 关键词: DNA; analytical method; birefringences; copolymer I; electrophoresis; gel electrophoresis; light scattering; low angle X ray diffraction analysis; method development; micelles; nonblood rheology; nuclear magnetic resonance spectroscopy; nucleic acid sequence; polymerization; solutions; viscosity

In DNA sequencing analysis by means of electrophoresis, the separation medium could use further improvements. The long-term objective of the proposed project is to develop a better separation medium 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 central theme is to take advantage of the self-assembly behavior of block copolymers so that predesigned polymer networks can be formulated by considering the chemical nature of the blocks, the total chain length, the block length ratio and the molecular architecture of the blocks in a selective solvent which is the buffer solution used in DNA sequencing analysis. In the self-assembled polymer networks, it is expected that several desirable properties can be designed into the properties of the separation medium. For examples, (1) at temperatures outside the operating temperature range for electrophoresis, the low molecular weight block copolymer (e.g. Pluronic polyols) solution has a low viscosity for ease of filling in capillary tubings or thin slabs, and (2) at the operating temperatures for electrophoresis, the self-assembled supramolecules can form gels of predetermined structures suitable for DNA sequencing analysis. In order to provide a basis for the design of a better separation medium, the structures of these entangled polymer chains (or gels) will be studied by means of physical techniques, such as viscosimetry, rheometry, laser light scattering, small angle neutron scattering, small angle x-ray scattering, C13 and proton NMR, and be correlated with electrophoresis results. By using the same physical techniques, it is proposed to study the effects of polydispersity on polymer chain entanglements and the use of a mixed polymer (e.g. polyacrylamide) solution/agarose gel approach which will be used to examine whether larger mesh size polymer networks can be achieved in order to resolve larger size DNA fragments. The agarose gel is used to restrict the translational motions of polyacrylamide chains so as to control the viscosity of the separation medium and the polymer chain dynamics.

在通过电泳的DNA测序分析中，分离 培养基可以使用进一步的改进。 长期目标 拟议的项目是开发更好的分离媒介 克服与当今聚丙烯酰胺有关的许多问题 溶液或凝胶：聚合物溶液通常太粘稠而无法填充 毛细管；聚合原位方法并非如此 可重现；聚合物凝胶的保质期有限。中心主题是 利用块共聚物的自组装行为， 可以通过考虑 块的化学性质，总链长，块长度 选择性溶剂中块的比率和分子结构 这是DNA测序分析中使用的缓冲溶液。 在 自组装的聚合物网络，预计会有几个理想的 可以在分离介质的性质中设计特性。 例如，（1）在工作温度范围以外的温度下 对于电泳，低分子量块共聚物（例如 pluronic polyols）溶液的粘度低，以便于填充 毛细管或薄板，（2）在工作温度下 电泳，自组装的超分子可以形成凝胶 预定的结构适合DNA测序分析。 为了为设计更好的分离介质的设计提供基础 将研究这些纠缠的聚合物链（或凝胶）的结构 通过物理技术，例如粘度法，风化计，激光 光散射，小角度中子散射，小角度X射线 散射，C13和质子NMR，与电泳相关 结果。 通过使用相同的物理技术，建议研究效果 在聚合物链纠缠上的多分散性和混合的使用 聚合物（例如聚丙烯酰胺）溶液/琼脂糖凝胶方法将是 用于检查是否可以实现较大的网格尺寸聚合物网络 为了解决较大尺寸的DNA片段。琼脂糖凝胶习惯 限制聚丙烯酰胺链的翻译动作，以便 控制分离介质和聚合物链的粘度 动力学。