Huang Ming-Feng, Huang Chih-Ching, Chang Huan-Tsung
Department of Chemistry, National Taiwan University, Taipei, Taiwan, ROC.
Electrophoresis. 2003 Sep;24(17):2896-902. doi: 10.1002/elps.200305432.
The analysis of double-stranded (ds) DNA fragments by capillary electrophoresis (CE) using poly(ethylene oxide) (PEO) solution containing gold nanoparticles (GNPs) is presented, focusing on evaluating size dependence of the GNPs and PEO on resolution and speed. To prevent the interaction of the capillary wall with DNA, the capillary was dynamically coated with polyvinylpyrrolidone. Using different PEO solutions containing GNPs ranging in diameter from 3.5 to 56 nm, we have achieved reproducible, rapid, and high-resolution DNA separations. The results indicate that the sizes of PEO and GNPs as well as the concentration of PEO affect resolution. The separation of DNA ranging in size from 8 to 2176 base pairs (bp) was accomplished in 5 min using 0.2% PEO (8 MDa) containing 56 nm GNPs. We have also demonstrated the separations of the DNA fragments ranging from 5 to 40 kbp using 0.05% PEO (2 MDa) containing 13 nm GNPs or 0.05% PEO (4 MDa) containing 32 nm GNPs. With very low viscosity (< 15 cP), automatic replacement of the sieving matrices is easy, indicating a great potential for high-throughput DNA analysis using capillary array electrophoresis systems.
本文介绍了使用含有金纳米颗粒（GNP）的聚环氧乙烷（PEO）溶液通过毛细管电泳（CE）分析双链（ds）DNA片段的方法，重点评估了GNP和PEO的尺寸对分辨率和速度的影响。为防止毛细管内壁与DNA相互作用，毛细管采用聚乙烯吡咯烷酮进行动态涂层处理。使用含有直径范围为3.5至56 nm的GNP的不同PEO溶液，我们实现了可重复、快速且高分辨率的DNA分离。结果表明，PEO和GNP的尺寸以及PEO的浓度会影响分辨率。使用含有56 nm GNP的0.2% PEO（8 MDa），在5分钟内完成了大小从8至2176碱基对（bp）的DNA分离。我们还展示了使用含有13 nm GNP的0.05% PEO（2 MDa）或含有32 nm GNP的0.05% PEO（4 MDa）对大小从5至40 kbp的DNA片段进行分离。由于粘度极低（< 15 cP），筛分基质易于自动更换，这表明使用毛细管阵列电泳系统进行高通量DNA分析具有巨大潜力。
