Nature Biotechnology
17, 365 - 370 (1999)
doi:10.1038/7921
Single nucleotide polymorphic discrimination by an electronic dot blot
assay on semiconductor microchipsPatrick N. Gilles1, 3, David J. Wu1, Charles B. Foster2, Patrick J. Dillon1, 3
& Stephen J. Chanock21
Nanogen Inc., 10398 Pacific Center Ct.
, San Diego, CA, 92121. 2
Pediatric Oncology Branch, National Cancer Institutes,
National Institute of Health, 9000 Rockville Pike,
Bethesda, MD, 20892. 3
Present address, Invitrogen Corporation,
1600 Faraday, Carlsbad, CA, 92008.
Correspondence should be addressed to David J. Wu dwu@nanogen.com).single nucleotide polymorphismmannose binding proteinmicroarraychipWe have developed a rapid assay for single nucleotide polymorphism (SNP)
detection that utilizes electronic circuitry on silicon microchips. The method
was validated by the accurate discrimination of blinded DNA samples for the
complex quadra-allelic SNP of mannose binding protein. The microchip directed
the transport, concentration, and attachment of amplified patient DNA to selected
electrodes (test sites) creating an array of DNA samples. Through control
of the electric field, the microchip enabled accurate genetic identification
of these samples using fluorescently labeled DNA reporter probes. The accuracy
of this approach was established by internal controls of dual labeled reporters
and by using mismatched sequences in addition to the wild-type and variant
reporter sequences to validate the SNP-genotype. The ability to customize
this assay for multiple genes has advantages over other existing approaches.
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