有利于多硼酸盐和路易斯酸配合物形成的条件下改进的 DNA 电泳。

Improved DNA electrophoresis in conditions favoring polyborates and lewis acid complexation.

机构信息

Department of Biomedical Engineering, The Johns Hopkins University and the Sidney Kimmel Comprehensive Center at Johns Hopkins, Baltimore, Maryland, United States of America.

出版信息

PLoS One. 2010 Jun 25;5(6):e11318. doi: 10.1371/journal.pone.0011318.

DOI:10.1371/journal.pone.0011318

PMID:20593002

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2892876/

Abstract

Spatial compression among the longer DNA fragments occurs during DNA electrophoresis in agarose and non-agarose gels when using certain ions in the conductive buffer, impairing the range of fragment sizes resolved well in a single gel. Substitutions using various polyhydroxyl anions supported the underlying phenomenon as the complexation of Lewis acids to DNA. We saw significant improvements using conditions (lithium borate 10 mM cations, pH 6.5) favoring the formation of borate polyanions and having lower conductance and Joule heating, delayed electrolyte exhaustion, faster electrophoretic run-speed, and sharper separation of DNA bands from 100 bp to 12 kb in a single run.

摘要

在琼脂糖和非琼脂糖凝胶中进行 DNA 电泳时，如果在导电缓冲液中使用某些离子，较长的 DNA 片段之间会发生空间压缩，从而影响在单个凝胶中良好分辨的片段大小范围。使用各种多羟基阴离子进行取代支持了这一现象，因为路易斯酸与 DNA 的络合作用。我们发现使用有利于形成硼酸多阴离子的条件（10mM 锂硼酸盐阳离子，pH6.5）有显著的改进，这些条件具有较低的电导率和焦耳加热、延迟电解质耗尽、更快的电泳运行速度以及在单次运行中从 100bp 到 12kb 的 DNA 带的更清晰分离。