title: Genotyping of mutations in the beta-globin gene using allele specific hybridization.
abstract: The use of DNA microarrays for genotyping is economically favorable when compared to real-time PCR and DNA sequencing. Here, we demonstrate a DNA microarray-based assay using allele-specific oligonucleotide (ASO) probes for genotyping mutations in the beta-globin gene. The assay makes use of agarose film coated glass slides as substrate, unmodified melting temperature matched ASO probes, target amplification and labeling using T7 in vitro transcription, mixing during hybridization and finally visualization using a fluorescent scanner. In this chapter we will emphasize on probe design, optimization, and validation of an allele-specific hybridization assay.
authors.affiliation: Technical University of Denmark, Kgs, Lyngby, Denmark.
authors.forename: Lena
authors.initials: L
authors.lastname: Poulsen
authors.forename: Jesper
authors.initials: J
authors.lastname: Petersen
authors.forename: Martin
authors.initials: M
authors.lastname: Dufva
mesh_terms: D000483:Alleles; D015342:DNA Probes / Q000378:metabolism; D005838:Genotype; D006801:Humans; D009154:Mutation / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012685:Sepharose; D055544:beta-Globins / Q000235:genetics*
chemical_list: D015342:DNA Probes; D055544:beta-Globins; D012685:Sepharose