Factors to be considered for robust high-throughput automated DNA sequencing using a multiple-capillary array instrument

Emanuel Carrilho; Arthur W. Miller; Marie C. Ruiz-Martinez; Lev Kotler; Jeffrey Kesilman; Barry L. Karger

doi:10.1117/12.274344

22 May 1997 Factors to be considered for robust high-throughput automated DNA sequencing using a multiple-capillary array instrument

Emanuel Carrilho, Arthur W. Miller, Marie C. Ruiz-Martinez, Lev Kotler, Jeffrey Kesilman, Barry L. Karger

Author Affiliations +

Proceedings Volume 2985, Ultrasensitive Biochemical Diagnostics II; (1997) https://doi.org/10.1117/12.274344
Event: BiOS '97, Part of Photonics West, 1997, San Jose, CA, United States

Abstract

The overall goal of our program is to develop a robust, high throughput, fully automated DNA sequencing instrument based on replaceable polymer solutions using a multicapillary array. Significant effort has already been devoted to column and polymer chemistry in order to obtain long read lengths per run in fast analysis time. In this paper we report on progress in instrument considerations and data processing software. A simple instrument design, based on no moving parts for continuous illumination of the capillaries and detection of the fluorescent light was used for this study. Our polymer solution replacement system with the permanent connection between the buffer/chamber manifold and capillary columns on the detector side is designed to prevent the trapping of air bubbles during matrix solution replacement. A special construction of a column-electrode couple on the injection side precludes air trapping during sample injection from small sample volumes. Our in-house software now features the significant reduction of the crosstalk signal from neighbor columns, which may be a potential problem in densely packed large capillary array sequencers.

Citation Download Citation

Emanuel Carrilho, Arthur W. Miller, Marie C. Ruiz-Martinez, Lev Kotler, Jeffrey Kesilman, and Barry L. Karger "Factors to be considered for robust high-throughput automated DNA sequencing using a multiple-capillary array instrument", Proc. SPIE 2985, Ultrasensitive Biochemical Diagnostics II, (22 May 1997); https://doi.org/10.1117/12.274344

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