Effect of substrate thermal resistance on space-domain microchannel fluorescent melting curve analysis

David J. Kinahan, Tara M. Dalton, Mark R.D. Davies

Research output: Contribution to journalArticlepeer-review

Abstract

In recent years, Fluorescent Melting Curve Analysis (FMCA) has become an almost ubiquitous feature of commercial quantitative PCR (qPCR) thermal cyclers. Here a micro-fluidic device is presented capable of performing FMCA within a microchannel. The device consists of modular thermally conductive blocks which can sandwich a microfluidic substrate. Opposing ends of the blocks are held at differing temperatures and a linear thermal gradient is generated along the microfluidic channel. Fluorescent measurements taken from a sample as it passes along the micro-fluidic channel permits fluorescent melting curves to be generated. In this study we measure DNA melting temperature from two plasmid fragments. The effects of flow velocity and ramp-rate are investigated, and measured melting curves are compared to those acquired from a commercially available PCR thermocycler.

Original languageEnglish
Pages (from-to)747-754
Number of pages8
JournalBiomedical microdevices
Volume11
Issue number4
DOIs
Publication statusPublished - 2009

Keywords

  • μTAS
  • DNA denaturation
  • Fluorescent melting curve analysis
  • Microchannel
  • PCR
  • Thermal resistance

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