Fast capillary-scanning system for detecting fluorescently labelled DNA sequencing fragments separated by capillary gel electrophoresis
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Summary
A capillary-scanning system for increasing the total throughput of DNA sequencing has been developed. The DNA sequencing method using the system features a spatial and temporal separation of laser-induced fluorescence detection from capillary gel electrophoresis. Fluorescently labelled adenine base fragments of pBluescript SK(−) were, produced in enzymatic sequencing reactions, and separated by capillary gel electrophoresis using UV-visible transparent capillaries filled with 8% T, 0% C polyacrylamide gel. The capillary containing all bands of the fragments was then scanned longitudinally with a 488 nm argon ion laser beam in 2.6 min. The adenine base sequence of the DNA was determined out to 400 bases by detecting fluorescence signals generated from the bands during the scan. The present scan speed is essentially limited by a slow strip-chart recorder and could be greatly increased by employing a fast data acquisition system.
Key Words
Capillary gel electrophoresis Laser-induced fluorescence DNA sequencing Capillary-scanningPreview
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