Beijing National Laboratory for Molecular Sciences (BNLMS), College of Chemistry, Peking University, Beijing, China.
J Chromatogr A. 2011 May 20;1218(20):3037-41. doi: 10.1016/j.chroma.2011.03.027. Epub 2011 Mar 21.
The separation and sequencing of DNA are the main objectives of the Human Genome Project, and this project has also been very useful for gene analysis and disease diagnosis. Capillary electrophoresis (CE) is one of the most common techniques for the separation and analysis of DNA. DNA separations are usually achieved using capillary gel electrophoresis (CGE) mode, in which polymer gel is packed into the capillary. Compared with a traditional CGE matrix, a hydrophilic polymer matrix, which can be adsorb by the capillary wall has numerous advantages, including stability, reproducibility and ease of automation. Various water-soluble additives, such as linear poly(acrylamide) (PAA) and poly(N,N-dimethylacrylamide) (PDMA), have been employed as media. In this study, different star-shaped PDMA polymers were designed and synthesized to achieve lower polymer solution viscosity. DNA separations with these polymers avoid the disadvantages of high viscosity and long separation time while maintaining high resolution (10 bp between 271 bp and 281 bp). The influences of the polymer concentration and structure on DNA separation were also determined in this study; higher polymer concentration yielded better separation performance, and star-like polymers were superior to linear polymers. This work indicates that modification of the polymer structure is a potential strategy for optimizing DNA separation.
DNA 的分离和测序是人类基因组计划的主要目标,该计划也非常有助于基因分析和疾病诊断。毛细管电泳(CE)是分离和分析 DNA 的最常用技术之一。DNA 分离通常采用毛细管凝胶电泳(CGE)模式,在该模式中聚合物凝胶被填充到毛细管中。与传统的 CGE 基质相比,亲水聚合物基质可以被毛细管壁吸附,具有许多优点,包括稳定性、重现性和易于自动化。线性聚(丙烯酰胺)(PAA)和聚(N,N-二甲基丙烯酰胺)(PDMA)等各种水溶性添加剂已被用作介质。在这项研究中,设计并合成了不同的星形 PDMA 聚合物,以实现较低的聚合物溶液粘度。使用这些聚合物进行 DNA 分离可以避免高粘度和长分离时间的缺点,同时保持高分辨率(271bp 和 281bp 之间的 10bp)。本研究还确定了聚合物浓度和结构对 DNA 分离的影响;较高的聚合物浓度可获得更好的分离性能,且星形聚合物优于线性聚合物。这项工作表明,聚合物结构的修饰是优化 DNA 分离的一种潜在策略。