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

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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 10{sup 6} smaller than commercial real-time PCR systems. 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 thermal cycled through the PCR protocol without droplet motion. With this system a 10-pL droplet, encapsulating less than one copy of viral genomic ... continued below

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PDF-file: 19 pages; size: 3.1 Mbytes

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Beer, N R; Hindson, B; Wheeler, E; Hall, S B; Rose, K A; Kennedy, I et al. April 20, 2007.

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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 10{sup 6} smaller than commercial real-time PCR systems. 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 thermal 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 {approx}18, twenty 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.

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PDF-file: 19 pages; size: 3.1 Mbytes

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  • Journal Name: Analytical Chemistry, vol. 79, no. 22, November 15, 2007, pp. 8471-8475; Journal Volume: 79; Journal Issue: 22

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  • Report No.: UCRL-JRNL-230233
  • Grant Number: W-7405-ENG-48
  • Office of Scientific & Technical Information Report Number: 929191
  • Archival Resource Key: ark:/67531/metadc898235

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Office of Scientific & Technical Information Technical Reports

Reports, articles and other documents harvested from the Office of Scientific and Technical Information.

Office of Scientific and Technical Information (OSTI) is the Department of Energy (DOE) office that collects, preserves, and disseminates DOE-sponsored research and development (R&D) results that are the outcomes of R&D projects or other funded activities at DOE labs and facilities nationwide and grantees at universities and other institutions.

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  • April 20, 2007

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  • Sept. 27, 2016, 1:39 a.m.

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  • Nov. 30, 2016, 2:07 p.m.

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Beer, N R; Hindson, B; Wheeler, E; Hall, S B; Rose, K A; Kennedy, I et al. On-chip real-time single-copy polymerase chain reaction in picoliter droplets, article, April 20, 2007; Livermore, California. (digital.library.unt.edu/ark:/67531/metadc898235/: accessed November 23, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.