MODEL AND COMPUTER-SIMULATIONS OF THE MOTION OF DNA-MOLECULES DURING PULSE FIELD GEL-ELECTROPHORESIS

被引:33
作者
SMITH, SB [1 ]
HELLER, C [1 ]
BUSTAMANTE, C [1 ]
机构
[1] UNIV CONSTANCE, FAK BIOL, W-7750 CONSTANCE, GERMANY
关键词
D O I
10.1021/bi00235a021
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A model is presented for the motion of individual molecules of DNA undergoing pulse field gel electrophoresis (PFGE). The molecule is represented by a chain of charged beads connected by entropic springs, and the gel is represented by a segmented tube surrounding the beads. This model differs from earlier reptation/tube models in that the tube is allowed to leak in certain places and the chain can double over and flow out of the side of the tube in kinks. It is found that these kinks often lead to the formation of U shapes, which are a major source of retardation in PFGE. The results of computer simulations using this model are compared with real DNA experimental results for the following cases: steady field motion as seen in fluorescence microscopy, mobility in steady fields, mobility in transverse field alternation gel electrophoresis (TFAGE), mobility in field inversion gel electrophoresis (FIGE), and linear dichroism (LD) of DNA in agarose gels during PFGE. Good agreement between the simulations and the experimental results is obtained.
引用
收藏
页码:5264 / 5274
页数:11
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