聚丙烯酰胺
回转半径
DNA
丙烯酰胺
化学
单体
聚丙烯酰胺凝胶电泳
半径
分析化学(期刊)
凝胶电泳
分子
材料科学
聚合物
高分子化学
色谱法
有机化学
生物化学
计算机安全
计算机科学
酶
作者
Diana L. Holmes,Nancy Stellwagen
标识
DOI:10.1002/elps.1150120405
摘要
Abstract The mobilities of normal and anomalously migrating DNA fragments were determined in polyacrylamide gels of different acrylamide concentrations, polymerized with 3 % N , N ′‐methylenebisacrylamide as the crosslinker. The DNA samples were a commercially available 123‐bp ladder and two molecular weight ladders containing multiple copies of two 147‐base pair (bp) restriction fragments, obtained from the MspI digestion of plasmid pBR322. One of the 147 bp fragments is known to migrate anomalously slowly in polyacrylamide gels. Ferguson plots were constructed for all multimer ladders, using both absolute mobilities and relative mobilities with respect to the smallest DNA molecule in each data set. If the retardation coefficients were calculated from the relative mobilities, and the rms radius of gyration was used as the measure of DNA size, the Ogston equations were obeyed and the gel fiber parameters could be calculated. The effective pore sizes of the gels were estimated from the gel concentration at which the mobility of a given DNA molecule was reduced to one‐half its mobility at zero gel concentration. The estimated pore radii ranged from ∼ 130 nm for 3.5 % gels to ∼ 70 nm for 10.5 % gels. These values are much larger than the pore sizes previously determined for the polyacrylamide matrix.
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