High-speed DNA genotyping using microfabricated capillary array electrophoresis chips.
Anal Chem
; 69(11): 2181-6, 1997 Jun 01.
Article
em En
| MEDLINE
| ID: mdl-9183181
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 bis-intercalation 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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Antígenos de Histocompatibilidade Classe I
/
Eletroforese Capilar
/
Hemocromatose
/
Antígenos HLA
/
Proteínas de Membrana
Limite:
Humans
Idioma:
En
Revista:
Anal Chem
Ano de publicação:
1997
Tipo de documento:
Article