On-chip, real-time, single-copy polymerase chain reaction in picoliter droplets

N. Reginald Beer, Benjamin J. Hindson, Elizabeth K. Wheeler, Sara B. Hall, Klint A. Rose, Ian M. Kennedy, Bill W. Colston

Research output: Contribution to journalArticle

243 Scopus citations

Abstract

The first lab-on-chip system for picoliter droplet generation and PCR amplification with real-time fluorescence detection has performed PCR in isolated droplets at volumes 106 smaller than commercial real-time PCR instruments. The system utilized a shearing T-junction in a silicon device to generate a stream of monodisperse picoliter droplets that were isolated from the microfluidic channel walls and each other by the oil-phase carrier. An off-chip valving system stopped the droplets on-chip, allowing them to be thermally cycled through the PCR protocol without droplet motion. With this system, a 10-pL droplet, encapsulating less than one copy of viral genomic DNA through Poisson statistics, showed real-time PCR amplification curves with a cycle threshold of ∼18, 20 cycles earlier than commercial instruments. This combination of the established real-time PCR assay with digital microfluidics is ideal for isolating single-copy nucleic acids in a complex environment.

Original languageEnglish (US)
Pages (from-to)8471-8475
Number of pages5
JournalAnalytical Chemistry
Volume79
Issue number22
DOIs
Publication statusPublished - Nov 15 2007

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ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Beer, N. R., Hindson, B. J., Wheeler, E. K., Hall, S. B., Rose, K. A., Kennedy, I. M., & Colston, B. W. (2007). On-chip, real-time, single-copy polymerase chain reaction in picoliter droplets. Analytical Chemistry, 79(22), 8471-8475. https://doi.org/10.1021/ac701809w