Tabuchi Mari, Katsuyama Yoshinori, Nogami Kazuma, Nagata Hideya, Wakuda Keisuke, Fujimoto Masayuki, Nagasaki Yukio, Yoshikawa Kenichi, Kataoka Kazunori, Baba Yoshinobu
Department of Molecular and Pharmaceutical Biotechnology, Graduate School of Pharmaceutical Sciences, The University of Tokushima, CREST, JST, 21st COE, Tokushima 770-8505, Japan.
Lab Chip. 2005 Feb;5(2):199-204. doi: 10.1039/b410498f. Epub 2004 Dec 16.
We report here advanced microchip electrophoresis using a nanoparticle doped polymer solution that enables greater separation of DNA. The proposed system is simple and effective without any new apparatus or complicated procedures. Various amounts and sizes (80 nm, 110 nm, and 193 nm) of polymer nanoparticle solutions (PEGylated-latex) were mixed with a conventional polymer solution for microchip electrophoresis. When a 0.49 wt% hydroxyl propyl methyl cellulose (HPMC) buffer solution was mixed with a 2.25 wt% 80 nm-PEGylated-latex a higher separation efficiency and a higher mobility of a wider molecular range of dsDNA (10 bp to 2 kbp) was achieved under low viscosity conditions (<5.5 cP) than in conventional 0.7% HPMC. The separation performance was dependent on the particle size and concentration. Furthermore, the effectiveness of the larger PEGylated-latex (193 nm) was not as high as the smaller one (80 to 110 nm). The observed separation improvement by polymer solution with latex-nanoparticles seems to derive from the balance between wider polymer mesh size and the structural obstacles of particles in the buffer.
我们在此报告一种先进的微芯片电泳技术,该技术使用掺杂纳米颗粒的聚合物溶液,能够实现对DNA更好的分离。所提出的系统简单有效,无需任何新设备或复杂程序。将不同数量和尺寸(80纳米、110纳米和193纳米)的聚合物纳米颗粒溶液(聚乙二醇化乳胶)与用于微芯片电泳的传统聚合物溶液混合。当将0.49重量%的羟丙基甲基纤维素(HPMC)缓冲溶液与2.25重量%的80纳米聚乙二醇化乳胶混合时,在低粘度条件下(<5.5厘泊),与传统的0.7% HPMC相比,双链DNA(10碱基对至2千碱基对)在更宽分子范围内实现了更高的分离效率和更高的迁移率。分离性能取决于颗粒大小和浓度。此外,较大的聚乙二醇化乳胶(193纳米)的效果不如较小的(80至110纳米)。观察到的含乳胶纳米颗粒的聚合物溶液对分离的改善似乎源于更宽的聚合物网孔尺寸与缓冲液中颗粒的结构障碍之间的平衡。