Fast DNA separations using poly(ethylene oxide) in non-denaturing medium with temperature programming

被引：25

作者：

Fung, EN ^{[1
]}

Pang, HM ^{[1
]}

Yeung, ES ^{[1
]}

机构：

[1] US DOE, Ames Lab, Ames, IA 50011 USA

来源：

JOURNAL OF CHROMATOGRAPHY A | 1998年 / 806卷 / 01期

关键词：

gel preparation; DNA; poly(ethylene oxide);

D O I：

10.1016/S0021-9673(97)00812-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We demonstrated fast DNA separations in low viscosity entangled solutions with a temperature gradient in a nondenaturing separation medium. The separations were carried out in a solution of commercially available poly(ethylene oxide) (PEO) [1xTris(hydroxymethyl)aminomethane berate buffer, without urea] with a temperature gradient of 2 degrees C/min. The performance was compared with that of a solution of PEO with urea at ambient temperature. We found that the former condition gives sufficient resolution for accurate base calling and that in general, it gave better separation for fragments larger than 450 base pairs (bp). Most importantly, the separation speed approaches 30 bp/min. In addition, we describe a simple yet reliable gel preparation protocol for such separations. (C) 1998 Elsevier Science B.V.

引用

收藏

页码：157 / 164

页数：8

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