Construction of a microfluidic chip, using dried-down reagents, for LATE-PCR amplification and detection of single-stranded DNA

Lab Chip. 2013 Dec 7;13(23):4635-41. doi: 10.1039/c3lc51049b.

Abstract

LATE-PCR is an advanced form of non-symmetric PCR that efficiently generates single-stranded DNA which can readily be characterized at the end of amplification by hybridization to low-temperature fluorescent probes. We demonstrate here for the first time that monoplex and duplex LATE-PCR amplification and probe target hybridization can be carried out in double layered PDMS microfluidics chips containing dried reagents. Addition of a set of reagents during dry down overcomes the common problem of single-stranded oligonucleotide binding to PDMS. These proof-of-principle results open the way to construction of inexpensive point-of-care devices that take full advantage of the analytical power of assays built using LATE-PCR and low-temperature probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Single-Stranded / analysis*
  • Dimethylpolysiloxanes / chemistry
  • Fluorescent Dyes / chemistry
  • Microfluidic Analytical Techniques / instrumentation*
  • Nucleic Acid Hybridization
  • Point-of-Care Systems
  • Polymerase Chain Reaction / instrumentation*
  • Temperature

Substances

  • DNA, Single-Stranded
  • Dimethylpolysiloxanes
  • Fluorescent Dyes
  • baysilon