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由聚(N-丙烯酰基-三(羟甲基)氨基甲烷)和聚乙烯吡咯烷酮形成的新型准互穿网络:用于通过紫外检测的毛细管电泳分离双链DNA和单链DNA片段的分离基质。

A new quasi-interpenetrating network formed by poly(N-acryloyl-tris-(hydroxymethyl)aminomethane and polyvinylpyrrolidone: separation matrix for double-stranded DNA and single-stranded DNA fragments by capillary electrophoresis with UV detection.

作者信息

Wang Qian, Xu Xu, Dai Lixin

机构信息

Laboratory of Analytical Chemistry, State Key Laboratory of Organometallic Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China.

出版信息

Electrophoresis. 2006 May;27(9):1749-57. doi: 10.1002/elps.200500597.

Abstract

The preparation of a new separation matrix, quasi-interpenetrating networks (quasi-IPNs) formed by poly(N-acryloyl-Tris) (poly(tris-A)) and PVP, and its application for dsDNA and ssDNA fragments separation by CE with UV detection, are presented. This new quasi-IPN exhibited high sieving performance, good dynamic coating ability, and low viscosity. Single-base resolutions of dsDNA fragments (Rs = 0.92 for 123/124 bp) and ssDNA fragments (Rs = 0.65 for 123/124 base, Rs = 0.48 for 309/310 base) were achieved by using the quasi-IPN of poly(tris-A)/PVP (2% + 2%) solution in a 31 cm effective length linear polyacrylamide (LPA)-coated column. Single-base separation of dsDNA fragments (Rs = 0.92 for 123/124 bp) was also obtained within 28 min in a 46.7 cm effective length bare column at higher 160 V/cm electric field strength by using the same quasi-IPN solution. The RSD of the migration time measured for each DNA fragments was less than 1.5% in the bare column for nine continuous runs. The effects of temperature and electric field strength on the DNA separation were also investigated.

摘要

本文介绍了一种新型分离基质——由聚(N-丙烯酰基三羟甲基氨基甲烷)(聚(三羟甲基氨基甲烷-A))和聚乙烯吡咯烷酮(PVP)形成的准互穿网络(quasi-IPN),及其在紫外检测毛细管电泳中用于双链DNA(dsDNA)和单链DNA(ssDNA)片段分离的应用。这种新型准互穿网络表现出高筛分性能、良好的动态涂层能力和低粘度。通过在31 cm有效长度的线性聚丙烯酰胺(LPA)涂层柱中使用聚(三羟甲基氨基甲烷-A)/PVP(2% + 2%)溶液的准互穿网络,实现了双链DNA片段(123/124 bp时分辨率Rs = 0.92)和单链DNA片段(123/124碱基时Rs = 0.65,309/310碱基时Rs = 0.48)的单碱基分辨率。在46.7 cm有效长度的裸柱中,使用相同的准互穿网络溶液,在160 V/cm的较高电场强度下,28分钟内也实现了双链DNA片段(123/124 bp时Rs = 0.92)的单碱基分离。在裸柱中连续九次运行时,每个DNA片段迁移时间的相对标准偏差(RSD)小于1.5%。还研究了温度和电场强度对DNA分离的影响。

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