Circulating polymerase chain reaction chips utilizing multiple-membrane activation

Chih Hao Wang, Yi Yu Chen, Chia Sheng Liao, Tsung Min Hsieh, Ching Hsing Luo, Jiunn Jong Wu, Huei Huang Lee, Gwo Bin Lee

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

This paper reports a new micromachined, circulating, polymerase chain reaction (PCR) chip for nucleic acid amplification. The PCR chip is comprised of a microthermal control module and a polydimethylsiloxane (PDMS)-based microfluidic control module. The microthermal control modules are formed with three individual heating and temperature-sensing sections, each modulating a specific set temperature for denaturation, annealing and extension processes, respectively. Micro-pneumatic valves and multiple-membrane activations are used to form the microfluidic control module to transport sample fluids through three reaction regions. Compared with other PCR chips, the new chip is more compact in size, requires less time for heating and cooling processes, and has the capability to randomly adjust time ratios and cycle numbers depending on the PCR process. Experimental results showed that detection genes for two pathogens, Streptococcus pyogenes (S. pyogenes, 777 bps) and Streptococcus pneumoniae (S. pneumoniae, 273 bps), can be successfully amplified using the new circulating PCR chip. The minimum number of thermal cycles to amplify the DNA-based S. pyogenes for slab gel electrophoresis is 20 cycles with an initial concentration of 42.5 pg νl-1. Experimental data also revealed that a high reproducibility up to 98% could be achieved if the initial template concentration of the S. pyogenes was higher than 4 pg νl-1.

Original languageEnglish
Article number024
Pages (from-to)367-375
Number of pages9
JournalJournal of Micromechanics and Microengineering
Volume17
Issue number2
DOIs
Publication statusPublished - 2007 Feb 1

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Mechanics of Materials
  • Mechanical Engineering
  • Electrical and Electronic Engineering

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