DNA sequencing using 96-capillary array electrophoresis.
Practical DNA sequencing in a rugged capillary array electrophoresis system coupled directly to 96-well microtiter plates is demonstrated. A CCD detector was used to monitor all capillaries simultaneously with laser-induced fluorescence at 1.75 frames per second. The reconstructed electropherograms show good signal-to-noise ratios and resolution for the entire capillary array. The system used standard dye labeling and image splitting to obtain fluorescence intensities in two wavelength regions to allow calling up to 410 bases for the DNA sequence. The use of a replaceable poly(ethylene oxide) matrix and a protective poly(vinylpyrrolidone) coating allows high separation speed and short turnaround time for high throughput DNA sequencing. Critical evaluation of the system performance over repeated runs with base calling is presented.
Pang
H
H
Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, 50011, USA.
Pavski
V
V
Yeung
E S
ES
D001331:Automation; D001483:Base Sequence; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D005453:Fluorescence; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*