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用于毛细管电泳高分辨率和高速分离DNA的聚(N-异丙基丙烯酰胺)-接枝-聚环氧乙烷

Poly(N-isopropylacrylamide)-g-poly(ethyleneoxide) for high resolution and high speed separation of DNA by capillary electrophoresis.

作者信息

Liang D, Song L, Zhou S, Zaitsev V S, Chu B

机构信息

Department of Chemistry, State University of New York at Stony Brook, 11794-3400, USA.

出版信息

Electrophoresis. 1999 Oct;20(14):2856-63. doi: 10.1002/(SICI)1522-2683(19991001)20:14<2856::AID-ELPS2856>3.0.CO;2-#.

Abstract

A new separation medium, poly(N-isopropylacrylamide)-g-poly(ethyleneoxide) (PNI-PAM-g-PEO) solution, used for double-stranded (ds) DNA separation by capillary electrophoresis (CE) is presented. This type of grafted copolymer has a good self-coating ability for quartz capillary tubing and a slightly temperature-dependent viscosity-adjustable property, making it easier to use. One bp resolution was achieved within 12.5 min by using 8% w/v PNIPAM-gPEO in 1 x TBE (Tris-borate-ethylenediaminetetraaceticacid) buffer with an effective column length of 10 cm and an applied electric field strength of 200 V/cm. The PNIPAM-g-PEO solutions had a high sieving ability for relatively small sized DNAs with the relative standard derivation for the first 10 runs being less than 0.9% by using the same polymer solution. With 8% w/v PNIPAM-g-PEO solution in a 1.5 cm column and 2400 V as the running voltage, phiX174/HaeIII digest could be clearly separated within 24 s.

摘要

介绍了一种用于毛细管电泳(CE)分离双链(ds)DNA的新型分离介质——聚(N-异丙基丙烯酰胺)-g-聚(环氧乙烷)(PNI-PAM-g-PEO)溶液。这种接枝共聚物对石英毛细管具有良好的自涂层能力,且粘度具有轻微的温度依赖性可调特性,使其更易于使用。在1×TBE(Tris-硼酸-乙二胺四乙酸)缓冲液中使用8%w/v的PNIPAM-gPEO,有效柱长为10 cm,施加电场强度为200 V/cm时,在12.5分钟内实现了1 bp的分辨率。PNIPAM-g-PEO溶液对相对较小尺寸的DNA具有较高的筛分能力,使用相同的聚合物溶液时,前10次运行的相对标准偏差小于0.9%。在1.5 cm柱中使用8%w/v的PNIPAM-g-PEO溶液,运行电压为2400 V时,phiX174/HaeIII酶切产物可在24秒内清晰分离。

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