High-speed DNA genotyping using microfabricated capillary array electrophoresis chips
- Univ. of California, Berkeley, CA (United States)
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.
- OSTI ID:
- 512893
- Journal Information:
- Analytical Chemistry (Washington), Journal Name: Analytical Chemistry (Washington) Journal Issue: 11 Vol. 69; ISSN ANCHAM; ISSN 0003-2700
- Country of Publication:
- United States
- Language:
- English
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