Hybridization of complementary strand and single-base mutated oligonucleotides detected with an on-line acoustic wave sensor.
The hybridization of a biotinylated 25-mer oligonucleotide probe with complementary, non-complementary and single-base mutated 25-mer targets at the liquid-solid (neutravidin-modified) interface of a thickness-shear mode acoustic wave device was studied. The sensor was incorporated into an on-line configuration capable of both variable flow and stop-flow experiments. Under ambient temperature conditions different signals were obtained for the complementary and non-complementary cases. At higher temperatures, the former system exhibits behaviour characteristic of the production of intermediate duplexes which are decomposed by the re-introduction of buffer solution. The use of these conditions allows the distinction of binding events involving a set of single-base mutated 25-mers. Different responses were obtained depending on both the nature of the instigated mismatch in base pairing and on the location of mutation.
Furtado
L M
LM
Department of Chemistry, University of Toronto, Ontario, Canada. bgranozi@alchemy.chem.utoronto.ca
Thompson
M
M
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D001330:Electronic Data Processing; D006801:Humans; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D017354:Point Mutation; D017422:Sequence Analysis, DNA; D013016:Sound
D015345:Oligonucleotide Probes