连续流动毛细管电泳中用于超灵敏DNA分析的荧光相关光谱法。

Fluorescence correlation spectroscopy for ultrasensitive DNA analysis in continuous flow capillary electrophoresis.

作者信息

Fogarty Keir, Van Orden Alan

机构信息

Department of Chemistry, Colorado State University, 1872 Campus Mail, Fort Collins, CO 80623, USA.

出版信息

Methods. 2009 Mar;47(3):151-8. doi: 10.1016/j.ymeth.2008.09.010. Epub 2008 Oct 11.

DOI:10.1016/j.ymeth.2008.09.010

PMID:18852049

Abstract

Continuous flow capillary electrophoresis (CFCE) is non-separations based analytical technique based on the free solution electrophoretic mobility of biological molecules such as DNA, RNA, peptides, and proteins. The electrophoretic mobilities and translational diffusion constants of the analyte molecules are determined using single molecule detection methods, including fluorescence correlation spectroscopy (FCS). CFCE is used to resolve multiple components in a mixture of analytes, measure electrophoretic mobility shifts due to binding interactions, and study the hydrodynamic and electrostatic properties of biological molecules in solution. Often this information is obtained with greater speed and sensitivity than conventational separations-based capillary-zone electrophoresis. This paper will focus on the application of two-beam fluorescence cross-correlation spectroscopy as a versatile detection method for CFCE and explore several applications to the study of the solution properties of single-stranded DNA.

摘要

连续流动毛细管电泳（CFCE）是一种基于生物分子（如DNA、RNA、肽和蛋白质）在自由溶液中的电泳迁移率的非分离分析技术。使用包括荧光相关光谱法（FCS）在内的单分子检测方法来测定分析物分子的电泳迁移率和平动扩散常数。CFCE用于分离分析物混合物中的多个组分，测量由于结合相互作用引起的电泳迁移率变化，并研究溶液中生物分子的流体动力学和静电性质。通常，与传统的基于分离的毛细管区带电泳相比，以更高的速度和灵敏度获得这些信息。本文将重点介绍双光束荧光交叉相关光谱法作为CFCE通用检测方法的应用，并探讨其在单链DNA溶液性质研究中的几种应用。