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用于寡核苷酸分离的纳米结构聚合物基质

Nanostructured polymer matrix for oligonucleotide separation.

作者信息

Zhang Jun, Burger Christian, Chu Benjamin

机构信息

Department of Chemistry, Stony Brook University, NY 11794-3400, USA.

出版信息

Electrophoresis. 2006 Sep;27(17):3391-8. doi: 10.1002/elps.200500839.

Abstract

A nanostructured copolymer matrix has successfully separated oligonucleotides with high resolution by CE using a very short separation channel which simulates the real microchip condition for the first time. The triblock copolymer, E(45)B(14)E(45) (B20-5000) with E, B, and subscript denoting oxyethylene, oxybutylene, and segment length, respectively, has a unique temperature-dependent viscosity-adjustable property and a dynamic coating ability in 1xTBE buffer (89 mM Tris, 89 mM boric acid, 2 mM EDTA in Milli-Q water). The B-block is hydrophilic at low temperatures, e.g., 4 degrees C, and the polymer solution has a very low viscosity of about 100 cP in a 32.5% w/v solution. At room temperatures, the B-block becomes hydrophobic due to a breakdown of hydrogen bonds between B-blocks and water, and the polymer matrix forms a body-centered cubic structure at high concentrations. Oligonucleotide sizing markers ranging from 8 to 32 base could be successfully separated with one-base resolution in a 1.5 cm long separation channel by E(45)B(14)E(45) in its gel-like state.

摘要

一种纳米结构共聚物基质首次通过使用非常短的分离通道的毛细管电泳成功地以高分辨率分离了寡核苷酸,该分离通道模拟了真实的微芯片条件。三嵌段共聚物E(45)B(14)E(45)(B20 - 5000),其中E、B及下标分别表示氧化乙烯、氧化丁烯和链段长度,在1xTBE缓冲液(89 mM Tris、89 mM硼酸、2 mM EDTA于超纯水中)中具有独特的温度依赖性粘度可调特性和动态涂层能力。B链段在低温下,例如4℃时是亲水的,在32.5% w/v的溶液中聚合物溶液的粘度非常低,约为100 cP。在室温下,由于B链段与水之间的氢键断裂,B链段变得疏水,并且在高浓度下聚合物基质形成体心立方结构。在1.5 cm长的分离通道中,处于凝胶状状态的E(45)B(14)E(45)能够以单碱基分辨率成功分离8至32个碱基的寡核苷酸大小标准物。

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