Channel glass-based detection of human short insertion/deletion polymorphisms by tandem hybridization.
The development and critical evaluation of new technologies for identifying genetic polymorphisms will rapidly accelerate the discovery and diagnosis of disease-related genes. We report a novel way for distinguishing a new class of human DNA polymorphisms, short insertion/deletion polymorphisms (indels). A sensor with cylindrical pores named channel glass in combination with tandem hybridization, which uses a 5'-fluorescent labeled stacking probe and microarray-based short allele-specific oligonucleotide (capture probe) was investigated. This methodology allows indels to be detected individually and rapidly with small quantities of target DNA. This establishes a reliable quantitative test. Approaches for simultaneously hybridizing different targets to arrayed probes, designed to detect various indels in parallel, were examined. Five markers were consistently detected in a single hybridization. Possible factors impeding the hybridization reaction process are discussed.
Betanzos-Cabrera
Gabriel
G
Area Académica de Nutrición, Instituto de Ciencias de la Salud, Universidad Autónoma del Estado de Hidalgo, Hidalgo 42000, Mexico. gbetanzo@uaeh.reduaeh.mx
Harker
Brent W
BW
Doktycz
Mitchel J
MJ
Weber
James L
JL
Beattie
Kenneth L
KL
D001483:Base Sequence; D017353:Gene Deletion; D005838:Genotype; D005898:Glass; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*