Genotyping of mutations by combination of in-tube hybridization and universal tag-microarray

Abstract

An assay for detecting gene fragments, including: amplifying gene fragments comprising single-nucleotide polymorphisms (SNPs) to form an initial amplification product; isolating single strand oligonucleotides of interest from the initial amplification product; forming a solution comprising the oligonucleotides of interest and reporter molecules, each reporter molecule having a first oligonucleotide domain configured to hybridize with a complementary oligonucleotide of interest, and a second oligonucleotide domain configured to hybridize with a complementary capture probe; hybridizing, in the solution, the oligonucleotides of interest with the reporter molecules that have complementary first domains; applying the solution to the surface of a microarray including an array of capture probes fixed to a microarray slide; capturing the oligonucleotides of interest on the microarray by hybridizing the second oligonucleotide domains of the reporter molecules with complementary capture probes of the microarray; and detecting the hybridized oligonucleotides of interest captured on the microarray.