Rapid discrimination of single-nucleotide mismatches using a microfluidic device with monolayered beads.
A microfluidic device incorporating monolayered beads is developed for the discrimination of single-nucleotide mismatches, based on the differential dissociation kinetics between perfect match (PM) and mismatched (MM) duplexes. The monolayered beads are used as solid support for the immobilization of oligonucleotide probes containing a single-base variation. Target oligonucleotides hybridize to the probes, forming either PM duplexes or MM duplexes containing a single mismatch. Optimization studies show that PM and MM duplexes are easily discriminated based on their dissociation but not hybridization kinetics under an optimized buffer composition of 100mM NaCl and 50% formamide. Detection of single-nucleotide polymorphism (SNP) using the device is demonstrated within 8 min using four probes containing all the possible single-base variants. The device can easily be modified to integrate multiplexed detection, making high-throughput SNP detection possible.
Ng
Johnson Kian-Kok
JK
Division of Environmental Science and Engineering, National University of Singapore, EA-07-23, 9 Engineering Drive 1, Singapore 117576, Singapore.
Feng
Hanhua
H
Liu
Wen-Tso
WT
D020137:Base Pair Mismatch; D001483:Base Sequence; D015342:DNA Probes; D044085:Microfluidics / Q000295:instrumentation*; D009711:Nucleotides / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide
D015342:DNA Probes; D009711:Nucleotides