Frank R, Köster H
Nucleic Acids Res. 1979;6(6):2069-87. doi: 10.1093/nar/6.6.2069.
The specific influence of the four nucleobases on electrophoretic mobility of oligodeoxyribonucleotides in polyacrylamide-gels under denaturing and nondenaturing conditions has been investigated using homooligomers from the four deoxyribonucleotides as chain length standards. Homooligomers of same chain lengths exhibit remarkable differences in mobility. Specific retardation of any other oligonucleotide investigated was found to be mainly dependent on base composition but not on sequence. A simple procedure is presented for calculating mobilities relative to the standards on denaturing gels. This allows a reliable identification of oligonucleotides on acrylamide-gels by exact chain length determination with respect to base composition and furthermore a detailed interpretation of complex reaction mixtures. The homooligomers also show the same differences in mobility on nondenaturing gels. The significance of this effect for strand separation is discussed.
使用来自四种脱氧核糖核苷酸的同聚物作为链长标准,研究了四种核碱基在变性和非变性条件下对聚丙烯酰胺凝胶中寡脱氧核糖核苷酸电泳迁移率的具体影响。相同链长的同聚物在迁移率上表现出显著差异。所研究的任何其他寡核苷酸的特定阻滞主要取决于碱基组成而非序列。本文提出了一种简单的方法来计算变性凝胶上相对于标准物的迁移率。这使得通过精确测定碱基组成的链长,能够在丙烯酰胺凝胶上可靠地鉴定寡核苷酸,进而对复杂反应混合物进行详细解释。同聚物在非变性凝胶上也表现出相同的迁移率差异。讨论了这种效应对于链分离的意义。