A comparison of hybridization efficiency between flat glass and channel glass solid supports.
Two different solid supports, channel glass and flat glass, were compared for their affect on the sensitivity and efficiency of DNA hybridization reactions. Both solid supports were tested using a set of arrayed, synthetic oligonucleotides that are designed to detect short insertion/deletion polymorphisms (SIDPs). A total of 13 different human SIDPs were chosen for analysis. Capture probes, designed for this test set, were covalently immobilized on substrates. Hybridization efficiency was assessed using fluorescently labeled stacking probes which were preannealed to the target and then hybridized to the support-bound oligonucleotide array; the hybridization pattern was detected by fluorescence imaging. It was found that structural features of nucleic acid capture probes tethered to a solid support and the molecular basis of their interaction with targets in solution have direct implications on the hybridization process. Our results demonstrate that channel glass has a number of practical advantages over flat glass including higher sensitivity and a faster hybridization rate.
Betanzos-Cabrera
Gabriel
G
Area Académica de Nutrición, Instituto de Ciencias de la Salud, Universidad Autónoma del Estado de Hidalgo, Abasolo 600 Pachuca de Soto, 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; D001709:Biotechnology; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D015342:DNA Probes / Q000235:genetics; D005456:Fluorescent Dyes; D005898:Glass; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA