DNA sequencing using four-color capillary array electrophoresis and energy-transfer primers

Indu Kheterpal; Jingyue Ju; Arun Radhakrishnan; Gabriel S. Brandt; Charles L. Ginther; Steven M. Clark; James J. Scherer; George F. Sensabaugh; Richard A. Mathies

doi:10.1117/12.237606

1 April 1996 DNA sequencing using four-color capillary array electrophoresis and energy-transfer primers

Indu Kheterpal, Jingyue Ju, Arun Radhakrishnan, Gabriel S. Brandt, Charles L. Ginther, Steven M. Clark, James J. Scherer, George F. Sensabaugh, Richard A. Mathies

Author Affiliations +

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237606
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

A practical capillary array electrophoresis sequencer has been constructed and tested for large scale DNA sequencing. This system employs a scanning confocal optical system to detect DNA fragments electrophoresed on a capillary array. By using the recently developed energy transfer fluorescent primers, sensitive four-color detection is facilitated while exciting with just a single laser wavelength (488 nm) from an argon ion laser. Software for facile reduction of the images to four-color trace files and for automated base-calling have also been developed. This system can detect up to 25 capillaries at a time and has a raw sequencing rate of approximately 6 kilobases/hour. Applications to mitochondrial D-loop DNA sequencing are presented as a demonstration.

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Indu Kheterpal, Jingyue Ju, Arun Radhakrishnan, Gabriel S. Brandt, Charles L. Ginther, Steven M. Clark, James J. Scherer, George F. Sensabaugh, and Richard A. Mathies "DNA sequencing using four-color capillary array electrophoresis and energy-transfer primers", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237606

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