Tan H V, Kitzis A, Berthollet T, Hamard G, Beldjord C, Benarous R
INSERM U15, Paris, France.
Nucleic Acids Res. 1988 Mar 25;16(5):1921-30. doi: 10.1093/nar/16.5.1921.
In contrast to all previous preparative electrophoresis apparatus which used a pump, electroendosmotic elution uses bound electrical charges at the end of the separating gel to generate a buffer flow. The electroendosmotic flow increased with increasing currents and decreasing buffer concentrations: its exact characteristics for the built apparatus were determined. The electroendosmotic device was able to separate two DNA fragments differing in size by only 5% with a recovery over 95%. As demonstrated in practical examples of recovery and uses of DNA inserts, up to 10 micrograms of DNA per band can be loaded at a time. The recovered DNA can be used directly for nick-translation, ligation... without further treatment. The performances of the method are expected to improve still further if the charge density and pores of the electroendosmotic medium can be "made-to-order" to provide a better flow profile of the eluting buffer.
与所有以前使用泵的制备电泳装置不同，电渗洗脱利用分离凝胶末端的结合电荷来产生缓冲液流。电渗流随着电流的增加和缓冲液浓度的降低而增加：确定了所构建装置的精确特性。该电渗装置能够分离大小仅相差5%的两个DNA片段，回收率超过95%。正如DNA插入片段回收和使用的实际例子所示，每条带一次可加载多达10微克的DNA。回收的DNA可直接用于缺口平移、连接……无需进一步处理。如果电渗介质的电荷密度和孔隙能够“定制”以提供更好的洗脱缓冲液流动曲线，预计该方法的性能将进一步提高。
