The limiting mobility of DNA sequencing fragments for both cross-linked and noncross-linked polymers in capillary electrophoresis: DNA sequencing at 1200 V cm(-1)

被引:35
作者
Yan, JY [1 ]
Best, N [1 ]
Zhang, JZ [1 ]
Ren, HJ [1 ]
Jiang, R [1 ]
Hou, J [1 ]
Dovichi, NJ [1 ]
机构
[1] UNIV ALBERTA,DEPT CHEM,EDMONTON,AB T6G 2G2,CANADA
关键词
capillary electrophoresis; DNA sequencing;
D O I
10.1002/elps.1150170611
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The mobility of DNA sequencing fragments was measured in Long-Ranger gels at an electric field ranging from 200 to 1200 V cm(-1) and in noncross-linked polyacrylamide at electric fields ranging from 100 to 300 V cm(-1). In both cases, N* the fragment length that denotes the onset of biased reptation with orientation, is inversely proportional to electric field. The inverse dependence of N* is inconsistent with the original biased reptation model but is consistent with modern models of DNA migration. While separation speed increases dramatically with electric field, the number of bases determined in a separation decreases in proportion to field strength. We present a DNA sequencing run at an electric field of 1200 V cm(-1). Roughly 200 bases of sequence are determined in 3.5 min.
引用
收藏
页码:1037 / 1045
页数:9
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