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Related Experiment Videos

High sensitivity PCR assay in plastic micro reactors.

Jianing Yang1, Yingjie Liu, Cory B Rauch

  • 1Physical Sciences Research Laboratories, Motorola Labs, Motorola, Inc, 7700 S. River Parkway, MD-ML34, Tempe, AZ 85284, USA.

Lab on a Chip
|April 22, 2004
PubMed
Summary

This study demonstrates a plastic micro PCR reactor

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Microfluidics

Background:

  • Micro PCR reactors offer rapid thermal cycling and small volume operation.
  • Assessing the sensitivity and specificity of micro PCR devices for rare target detection remains a challenge.

Purpose of the Study:

  • To design, construct, and evaluate a polycarbonate PCR micro reactor for rapid operation, efficiency, sensitivity, and specificity.
  • To investigate the detection limits and performance of the micro reactor for amplifying specific gene fragments.

Main Methods:

  • A serpentine-shaped, thin (0.75 mm) polycarbonate plastic PCR micro reactor was developed.
  • The reactor's performance was tested for amplifying a specific gene fragment of Escherichia coli (E. coli) K12.
  • Sensitivity was assessed at low template concentrations and in the presence of blood; specificity was evaluated using multiplex PCR.

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Main Results:

  • The micro reactor successfully amplified a 221 bp K12-specific gene product from as few as 10 E. coli cells in 30 minutes (30 cycles).
  • Detection sensitivity was confirmed by amplifying the target gene from 10 cells in 2% blood.
  • Specificity was demonstrated through multiplex PCR and amplification of different pathogen genes.

Conclusions:

  • This study presents the first systematic evaluation of assay sensitivity and specificity in plastic, disposable micro PCR devices.
  • The developed polycarbonate micro PCR reactor shows high sensitivity and specificity for detecting specific gene fragments.
  • These findings highlight the potential of microfluidic devices for sensitive and rapid molecular diagnostics.