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通过填充纳米颗粒的毛细管电泳分离长双链DNA。

Separation of long double-stranded DNA by nanoparticle-filled capillary electrophoresis.

作者信息

Huang Ming-Feng, Kuo Yi-Chun, Huang Chih-Ching, Chang Huan-Tsung

机构信息

Department of Chemistry, National Taiwan University, Section 4, Roosevelt Road, Taipei, Taiwan, ROC.

出版信息

Anal Chem. 2004 Jan 1;76(1):192-6. doi: 10.1021/ac034908u.

Abstract

We present the first example of the analysis of long double-stranded (ds) DNA molecules by nanoparticle-filled capillary electrophoresis (NFCE). To avoid aggregation of the gold nanoparticles (GNPs) and to allow strong interactions with the DNA molecules, the gold nanoparticles were modified with poly(ethylene oxide) (PEO) via noncovalent bonding to form gold nanoparticle/polymer composites (GNPPs). The neutral GNPPs are heavy (approximately 2.0 x 10(8) g/mol for the 32-nm GNP) and thus slow the DNA molecules that they encounter during the electrophoretic process. Compared to linear polymer solutions, such as hydroxyethyl cellulose and PEO, the GNPPs provide greater efficiency and require significantly shorter times to separate long dsDNA. The separation of lambda-DNA (0.12-23.1 kbp) by NFCE at -250 V/cm was accomplished in 3 min. The ability to separate high molecular weight DNA markers (8.27-48.5 kbp) with plate numbers greater than 10(6) suggests that this novel method may hold great promise for the analysis of long-stranded DNA molecules such as chromosomes. Moreover, this method is simple and affordable when compared to those that use micro- and nanofabricated devices for separating long DNA molecules.

摘要

我们展示了通过填充纳米颗粒的毛细管电泳(NFCE)分析长双链(ds)DNA分子的首个实例。为避免金纳米颗粒(GNP)聚集并确保其与DNA分子的强相互作用,通过非共价键用聚环氧乙烷(PEO)修饰金纳米颗粒,以形成金纳米颗粒/聚合物复合材料(GNPP)。中性的GNPP较重(对于32纳米的GNP,约为2.0×10⁸克/摩尔),因此会减缓其在电泳过程中遇到的DNA分子的移动速度。与线性聚合物溶液(如羟乙基纤维素和PEO)相比,GNPP具有更高的效率,分离长dsDNA所需的时间显著更短。在-250 V/cm下,通过NFCE在3分钟内完成了λ-DNA(0.12 - 23.1 kbp)的分离。能够以大于10⁶的塔板数分离高分子量DNA标志物(8.27 - 48.5 kbp)表明,这种新方法在分析诸如染色体等长链DNA分子方面可能具有巨大潜力。此外,与使用微纳制造设备分离长DNA分子的方法相比,该方法简单且成本低廉。

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