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.
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.
Yan
J Y
JY
Department of Chemistry, University of Alberta, Edmonton, Canada.
Best
N
N
Zhang
J Z
JZ
Ren
H J
HJ
Jiang
R
R
Hou
J
J
Dovichi
N J
NJ
D001483:Base Sequence; D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded / Q000032:analysis*; D004553:Electric Conductivity; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008969:Molecular Sequence Data; D011108:Polymers
D003432:Cross-Linking Reagents; D004277:DNA, Single-Stranded; D011108:Polymers