Mandecki W, Hayden M
Corporate Molecular Biology, Abbott Laboratories, Abbott Park, IL 60064.
DNA. 1988 Jan-Feb;7(1):57-62. doi: 10.1089/dna.1988.7.57.
We have found that 50 mM L-histidine pH 7.6 as a buffer for gel electrophoresis greatly improves the resolution of oligonucleotides less than 70 residues long on denaturing polyacrylamide gels. The histidine buffer increases spacing between DNA bands on the gel about twofold in comparison with a standard buffer (89 mM Tris-borate, 2 mM EDTA pH 8.3). In addition, low conductivity of the histidine buffer results in a threefold reduction of the electrophoresis time. Conditions for electrophoresis were optimized by varying both histidine and acrylamide concentrations. Other polycationic compounds, such as spermidine and ethylenediamine, were also tested for improved resolution of oligonucleotides. Several hypotheses as to the factors influencing the separation of DNA on gels are presented.
我们发现，50 mM pH 7.6的L-组氨酸作为凝胶电泳缓冲液，能显著提高变性聚丙烯酰胺凝胶上长度小于70个残基的寡核苷酸的分辨率。与标准缓冲液（89 mM Tris-硼酸，2 mM EDTA pH 8.3）相比，组氨酸缓冲液使凝胶上DNA条带之间的间距增加了约两倍。此外，组氨酸缓冲液的低电导率使电泳时间缩短了两倍。通过改变组氨酸和丙烯酰胺浓度对电泳条件进行了优化。还测试了其他聚阳离子化合物，如亚精胺和乙二胺，以提高寡核苷酸的分辨率。本文提出了几个关于影响凝胶上DNA分离的因素的假设。
