Improved separation of double-stranded DNA fragments by capillary electrophoresis using poly(ethylene oxide) solution containing colloids
Improved separation of double-stranded DNA fragments by capillary electrophoresis using poly(ethylene oxide) solution containing colloids
复制标题
DOI：
10.1002/elps.200305432
复制
发表时间：
2003-09-01
期刊：
影响因子：
2.9
通讯作者：
Chang, HT
中科院分区：
文献类型：
--
作者：
Huang, MF;Huang, CC;Chang, HT
采用毛细管电泳（CE）法，以聚环氧乙烷（PEO）为载金纳米粒子（GNP）的溶液，对双链DNA（ds）片段进行了分析，重点考察了GNP和PEO的大小对分辨率和速度的影响。为了防止毛细管壁与DNA的相互作用，用聚乙烯吡咯烷酮动态涂覆毛细管。使用不同的PEO解决方案含有GNP的直径范围从3.5到56 nm，我们已经实现了可重复的，快速的，高分辨率的DNA分离。结果表明，PEO和GNP的尺寸以及PEO的浓度影响分辨率。使用含有56nm GNP的0.2%PEO（8MDa）在5分钟内完成大小范围从8至2176个碱基对（bp）的DNA的分离。我们还证明了使用含有13 nm GNP的0.05%PEO（2 MDa）或含有32 nm GNP的0.05%PEO（4 MDa）分离5至40 kbp的DNA片段。由于粘度非常低（< 15 cP），筛分基质的自动更换很容易，这表明使用毛细管阵列电泳系统进行高通量DNA分析的巨大潜力。
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.