Three DNA sequencing methods using capillary gel electrophoresis and laser-induced fluorescence.
Capillary gel electrophoresis is demonstrated for the four-spectral-channel sequencing technique of Smith, the two-spectral-channel sequencing technique of Prober, and the one-spectral-channel sequencing technique of Richardson and Tabor. Sequencing rates up to 1000 bases/h are obtained at electric field strengths of 465 V/cm. At lower electric field strengths, capillary electrophoresis produces useful data for fragments greater than 550 nucleotides in length with 2 times better resolution than slab gel electrophoresis. An on-column detector produces detection limits of 200 zmol (1 zmol = 10(-21) mol = 600 molecules) for the four-spectral-channel technique. A postcolumn detector, based on the sheath flow cuvette, produces detection limits of 20 and 2 zmol for the two- and one-spectral-channel techniques, respectively.
Swerdlow
H
H
Department of Chemistry, University of Alberta, Edmonton, Canada.
Zhang
J Z
JZ
Chen
D Y
DY
Harke
H R
HR
Grey
R
R
Wu
S L
SL
Dovichi
N J
NJ
Fuller
C
C
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D004586:Electrophoresis; D005453:Fluorescence; D007834:Lasers
D004247:DNA