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一种用于通过微芯片电泳分离DNA的粘度可调聚合物。

A viscosity-tunable polymer for DNA separation by microchip electrophoresis.

作者信息

Kuroda Daisuke, Zhang Yong, Wang Jun, Kaji Noritada, Tokeshi Manabu, Baba Yoshinobu

机构信息

Department of Applied Chemistry, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.

出版信息

Anal Bioanal Chem. 2008 Aug;391(7):2543-9. doi: 10.1007/s00216-008-2196-4. Epub 2008 Jun 26.

Abstract

A thermo-responsive separation matrix, consisting of Pluronic F127 tri-block copolymers of poly(ethylene oxide) and poly(propylene oxide), was used to separate DNA fragments by microchip electrophoresis. At low temperature, the polymer matrix was low in viscosity and allowed rapid loading into a microchannel under low pressure. With increasing temperatures above 25 degrees C, the Pluronic F127 solution forms a liquid crystalline phase consisting of spherical micelles with diameters of 17-19 nm. The solution can be used to separate DNA fragments from 100 bp to 1500 bp on poly(methyl methacrylate) (PMMA) chips. This temperature-sensitive and viscosity-tunable polymer provided excellent resolution over a wide range of DNA sizes. Separation is based on a different mechanism compared with conventional matrices such as methylcellulose. To illustrate the separation mechanism of DNA in a Pluronic F127 solution, DNA molecular imaging was performed by fluorescence microscopy with F127 polymer as the separation matrix in microchip electrophoresis.

摘要

一种由聚环氧乙烷和聚环氧丙烷的普朗尼克F127三嵌段共聚物组成的热响应分离基质,被用于通过微芯片电泳分离DNA片段。在低温下,聚合物基质粘度较低,允许在低压下快速加载到微通道中。随着温度升高到25摄氏度以上,普朗尼克F127溶液形成由直径为17 - 19纳米的球形胶束组成的液晶相。该溶液可用于在聚甲基丙烯酸甲酯(PMMA)芯片上分离100 bp至1500 bp的DNA片段。这种对温度敏感且粘度可调的聚合物在广泛的DNA大小范围内提供了出色的分辨率。与传统基质如甲基纤维素相比,分离基于不同的机制。为了阐明DNA在普朗尼克F127溶液中的分离机制,在微芯片电泳中以F127聚合物作为分离基质,通过荧光显微镜进行了DNA分子成像。

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