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使用可变角度轮廓夹式均匀电场装置分离大型DNA

Separation of large DNA by a variable-angle contour-clamped homogeneous electric field apparatus.

作者信息

Chu G, Gunderson K

机构信息

Department of Medicine, Stanford University Medical Center, California 94305.

出版信息

Anal Biochem. 1991 May 1;194(2):439-46. doi: 10.1016/0003-2697(91)90254-q.

Abstract

A device for separating large DNA molecules by pulsed field electrophoresis is described. Based on the principles of contour-clamped homogeneous electric fields (CHEF), it uses feedback to clamp voltages in a square electrode array, which is compact and inexpensive to construct, adaptable to computer control, and reorients the electric field by arbitrary angles. To illustrate its capabilities, pulsed fields with reorientation angles ranging from 90 to 140 degrees were used to separate DNAs of 4.7 and 5.7 megabases by up to four band-widths in 20 h. The combination of accessible technology and complete control of the electric field should facilitate the search for ways to resolve even larger DNA.

摘要

描述了一种用于通过脉冲场电泳分离大型DNA分子的装置。基于轮廓钳制均匀电场(CHEF)原理,它利用反馈来钳制方形电极阵列中的电压,该阵列结构紧凑且制造成本低,适用于计算机控制,并能将电场以任意角度重新定向。为了说明其功能,使用重定向角度范围为90至140度的脉冲场在20小时内将4.7和5.7兆碱基的DNA分离多达四个带宽。可获取的技术与对电场的完全控制相结合,应有助于寻找解析甚至更大DNA的方法。

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