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Title: Simultaneous monitoring of DNA fragments separated by electrophoresis in a multiplexed array of 100 capillaries

Abstract

Various excitation schemes for distributing a laser beam to a large number of capillaries in an array are evaluated. This led to the construction of a multiplexed system for monitoring the electrophoresis of DNA fragments in 100 capillaries. The laser-excited fluorescence signals from each capillary are simultaneously recorded at the rate of 0.6 frame/s by a CCD camera. The reconstructed electropherograms show excellent reproducibility and minimal cross-talk. The system provides for two simultaneous excitation wavelengths so that it can be adapted for two-color, two-intensity DNA sequencing based on the commercial four-dye chemistry. Only 20 mW per laser line was employed. Further development of this system to accommodate up to 4096 independent sequencing channels at a time is discussed. 26 refs., 6 figs., 2 tabs.

Authors:
;  [1]
  1. Ames Lab., IA (United States)
Publication Date:
OSTI Identifier:
7265367
DOE Contract Number:  
W-7405-ENG-82
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry (Washington); (United States)
Additional Journal Information:
Journal Volume: 66:9; Journal ID: ISSN 0003-2700
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; DNA SEQUENCING; MONITORING; CAMERAS; CAPILLARIES; CHARGE-COUPLED DEVICES; DYES; ELECTROPHORESIS; EXCITATION; FLUORESCENCE; LASER RADIATION; SEPARATION PROCESSES; BLOOD VESSELS; BODY; CARDIOVASCULAR SYSTEM; ELECTROMAGNETIC RADIATION; ENERGY-LEVEL TRANSITIONS; LUMINESCENCE; ORGANS; RADIATIONS; SEMICONDUCTOR DEVICES; STRUCTURAL CHEMICAL ANALYSIS; 550400* - Genetics; 550200 - Biochemistry; 400105 - Separation Procedures

Citation Formats

Ueno, Kyoji, and Yeung, E S. Simultaneous monitoring of DNA fragments separated by electrophoresis in a multiplexed array of 100 capillaries. United States: N. p., 1994. Web. doi:10.1021/ac00081a010.
Ueno, Kyoji, & Yeung, E S. Simultaneous monitoring of DNA fragments separated by electrophoresis in a multiplexed array of 100 capillaries. United States. https://doi.org/10.1021/ac00081a010
Ueno, Kyoji, and Yeung, E S. Sun . "Simultaneous monitoring of DNA fragments separated by electrophoresis in a multiplexed array of 100 capillaries". United States. https://doi.org/10.1021/ac00081a010.
@article{osti_7265367,
title = {Simultaneous monitoring of DNA fragments separated by electrophoresis in a multiplexed array of 100 capillaries},
author = {Ueno, Kyoji and Yeung, E S},
abstractNote = {Various excitation schemes for distributing a laser beam to a large number of capillaries in an array are evaluated. This led to the construction of a multiplexed system for monitoring the electrophoresis of DNA fragments in 100 capillaries. The laser-excited fluorescence signals from each capillary are simultaneously recorded at the rate of 0.6 frame/s by a CCD camera. The reconstructed electropherograms show excellent reproducibility and minimal cross-talk. The system provides for two simultaneous excitation wavelengths so that it can be adapted for two-color, two-intensity DNA sequencing based on the commercial four-dye chemistry. Only 20 mW per laser line was employed. Further development of this system to accommodate up to 4096 independent sequencing channels at a time is discussed. 26 refs., 6 figs., 2 tabs.},
doi = {10.1021/ac00081a010},
url = {https://www.osti.gov/biblio/7265367}, journal = {Analytical Chemistry (Washington); (United States)},
issn = {0003-2700},
number = ,
volume = 66:9,
place = {United States},
year = {1994},
month = {5}
}