Mathew M K, Smith C L, Cantor C R
Department of Genetics and Development, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Biochemistry. 1988 Dec 27;27(26):9210-6. doi: 10.1021/bi00426a020.
Bacteriophage DNAs annealed into linear oligomeric concatemers were used to examine the quantitative pulsed-field gel electrophoretic behavior of different-sized DNAs as a function of electrical field strength and pulse time. Three zones of resolution are observed for increasingly larger DNAs. In the first two zones, the electrophoretic mobility decreases linearly with increasing DNA size. The separation in zone 2 is roughly twice that in zone 1. The largest DNA molecules do not resolve at all and migrate in a compression zone. Mobility in zone 1 increases linearly with the electric field strength and decreases with the inverse of the pulse time. The behavior of DNA in zone 2 is qualitatively similar. However, the effect of field strength and pulse time on the separations in each zone is quite different. The results for zone 1 are generally consistent with the predictions of several existing physical models of pulsed-field gel electrophoresis, but no model accounts for all of the observed behavior in the three zones.
将退火形成线性寡聚连接体的噬菌体DNA用于研究不同大小DNA在脉冲场凝胶电泳中的定量行为，该行为是电场强度和脉冲时间的函数。对于越来越大的DNA，观察到三个分辨率区域。在前两个区域中，电泳迁移率随DNA大小增加而线性降低。区域2中的分离度大约是区域1中的两倍。最大的DNA分子根本无法分辨，而是在压缩区迁移。区域1中的迁移率随电场强度线性增加，随脉冲时间的倒数降低。区域2中DNA的行为在定性上相似。然而，场强和脉冲时间对每个区域分离的影响有很大不同。区域1的结果通常与几种现有的脉冲场凝胶电泳物理模型的预测一致，但没有一个模型能解释三个区域中观察到的所有行为。
