Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchips.
We 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.
Gilles
P N
PN
Nanogen Inc., San Diego, CA 92121, USA.
Wu
D J
DJ
Foster
C B
CB
Dillon
P J
PJ
Chanock
S J
SJ
D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007375:Interleukin-1 / Q000235:genetics; D037241:Mannose-Binding Lectins; D008839:Microelectrodes; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes; D017354:Point Mutation; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D012666:Semiconductors
D002352:Carrier Proteins; D007375:Interleukin-1; D037241:Mannose-Binding Lectins; D015345:Oligonucleotide Probes