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Title: High-speed DNA genotyping using microfabricated capillary array electrophoresis chips

Abstract

Capillary array electrophoresis (CAE) chips have been designed and fabricated with the capacity to rapidly (<160 s) analyze 12 different samples in parallel. Detection of all lanes with 0.3 s temporal resolution was achieved using a laser-excited confocal-fluorescence scanner. The operation and capabilities of these CAE microdevices were first determined by performing electrophoretic separations of pBR322 MspI DNA samples. Genotyping of HLA-H, a candidate gene for the diagnosis of hereditary hemochromatosis, was then performed to demonstrate the rapid analysis of biologically relevant samples. Two-color multiplex fluorescence detection of HLA-H genotypes was accomplished by prelabeling the standard pBR322 MspI DNA ladder with a red emitting bisintercalation dye (butyl TOTIN) and on-column labeling of the HLA-H DNA with thiazole orange. This work establishes the feasibility of using CAE chips for high-speed, high-throughput genotyping. 44 refs., 7 figs.

Authors:
; ;  [1]
  1. Univ. of California, Berkeley, CA (United States)
Publication Date:
OSTI Identifier:
512893
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry (Washington)
Additional Journal Information:
Journal Volume: 69; Journal Issue: 11; Other Information: PBD: 1 Jun 1997
Country of Publication:
United States
Language:
English
Subject:
40 CHEMISTRY; 55 BIOLOGY AND MEDICINE, BASIC STUDIES; 42 ENGINEERING NOT INCLUDED IN OTHER CATEGORIES; DNA; SEPARATION PROCESSES; CHEMICAL ANALYSIS; LABELLING; ELECTROPHORESIS; CAPILLARIES; FLUORESCENCE; FABRICATION; DESIGN; LASERS; DYES; BIOTECHNOLOGY

Citation Formats

Woolley, A T, Sensabaugh, G F, and Mathies, R A. High-speed DNA genotyping using microfabricated capillary array electrophoresis chips. United States: N. p., 1997. Web. doi:10.1021/ac961237+.
Woolley, A T, Sensabaugh, G F, & Mathies, R A. High-speed DNA genotyping using microfabricated capillary array electrophoresis chips. United States. https://doi.org/10.1021/ac961237+
Woolley, A T, Sensabaugh, G F, and Mathies, R A. 1997. "High-speed DNA genotyping using microfabricated capillary array electrophoresis chips". United States. https://doi.org/10.1021/ac961237+.
@article{osti_512893,
title = {High-speed DNA genotyping using microfabricated capillary array electrophoresis chips},
author = {Woolley, A T and Sensabaugh, G F and Mathies, R A},
abstractNote = {Capillary array electrophoresis (CAE) chips have been designed and fabricated with the capacity to rapidly (<160 s) analyze 12 different samples in parallel. Detection of all lanes with 0.3 s temporal resolution was achieved using a laser-excited confocal-fluorescence scanner. The operation and capabilities of these CAE microdevices were first determined by performing electrophoretic separations of pBR322 MspI DNA samples. Genotyping of HLA-H, a candidate gene for the diagnosis of hereditary hemochromatosis, was then performed to demonstrate the rapid analysis of biologically relevant samples. Two-color multiplex fluorescence detection of HLA-H genotypes was accomplished by prelabeling the standard pBR322 MspI DNA ladder with a red emitting bisintercalation dye (butyl TOTIN) and on-column labeling of the HLA-H DNA with thiazole orange. This work establishes the feasibility of using CAE chips for high-speed, high-throughput genotyping. 44 refs., 7 figs.},
doi = {10.1021/ac961237+},
url = {https://www.osti.gov/biblio/512893}, journal = {Analytical Chemistry (Washington)},
number = 11,
volume = 69,
place = {United States},
year = {1997},
month = {6}
}