Kim Y, Morris M D
Department of Chemistry, University of Michigan, Ann Arbor 48109-1055.
Anal Chem. 1994 Apr 1;66(7):1168-74. doi: 10.1021/ac00079a035.
The effects of bis- and mono-intercalating dyes on the capillary electrophoretic separation of double-stranded DNA have been investigated in buffers containing (hydroxypropylmethyl)-cellulose. Broad bands and incomplete separations of phi X 174 DNA HaeIII digest were obtained with the bis-intercalators ethidium homodimer 1 (EthD-1) and ethidium homodimer 2 (EthD-2), under a wide range of dye and polymer concentrations. The above results were attributed to the presence of several intramolecular dye/DNA complexes for each fragment. For EthD-2/DNA complexes, but not for EthD-1/DNA complexes, additional bands beyond the expected number of fragments were observed. The extra bands were attributed to formation of intermolecular dimers. Use of the monomeric intercalating dyes, ethidium and propidium, allowed complete separation of all fragments of phi X 174 DNA HaeIII digest. A low-power green He-Ne laser (543.6 nm) was shown to be satisfactory for laser-induced fluorescence detection with any of these dyes.
在含有（羟丙基甲基）纤维素的缓冲液中，研究了双插入和单插入染料对双链DNA毛细管电泳分离的影响。在广泛的染料和聚合物浓度范围内，使用双插入剂乙锭同二聚体1（EthD-1）和乙锭同二聚体2（EthD-2）时，φX 174 DNA HaeIII酶切片段出现宽带和分离不完全的情况。上述结果归因于每个片段存在几种分子内染料/DNA复合物。对于EthD-2/DNA复合物，而非EthD-1/DNA复合物，观察到超出预期片段数目的额外条带。这些额外条带归因于分子间二聚体的形成。使用单体插入染料乙锭和碘化丙啶时，φX 174 DNA HaeIII酶切的所有片段都能完全分离。结果表明，低功率绿色氦氖激光器（543.6 nm）对于使用这些染料中的任何一种进行激光诱导荧光检测都令人满意。
