Rapid discrimination of single-nucleotide mismatches using a microfluidic device with monolayered beads

Johnson Kian Kok Ng, Hanhua Feng, Wen Tso Liu

Research output: Contribution to journalArticlepeer-review

Abstract

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 100 mM 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.

Original languageEnglish (US)
Pages (from-to)295-303
Number of pages9
JournalAnalytica Chimica Acta
Volume582
Issue number2
DOIs
StatePublished - Jan 23 2007
Externally publishedYes

Keywords

  • Dissociation kinetics
  • Microbeads
  • Microfluidic device
  • Single-nucleotide mismatches
  • Single-nucleotide polymorphisms (SNPs)

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy

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