On-chip single-copy real-time reverse-transcription PCR in isolated picoliter droplets

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The first lab-on-chip system for picoliter droplet generation and RNA isolation, followed by reverse transcription, and PCR amplification with real-time fluorescence detection in the trapped droplets has been developed. The system utilized a shearing T-junction in a fused silica device to generate a stream of monodisperse picoliter-scale 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 thermal cycling for reverse transcription and subsequent PCR amplification without droplet motion. This combination of the established real-time reverse transcription-PCR assay with digital microfluidics is ideal for ... continued below

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PDF-file: 21 pages; size: 2 Mbytes

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Beer, N R; Wheeler, E; Lee-Houghton, L; Watkins, N; Nasarabadi, S; Hebert, N et al. December 19, 2007.

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The first lab-on-chip system for picoliter droplet generation and RNA isolation, followed by reverse transcription, and PCR amplification with real-time fluorescence detection in the trapped droplets has been developed. The system utilized a shearing T-junction in a fused silica device to generate a stream of monodisperse picoliter-scale 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 thermal cycling for reverse transcription and subsequent PCR amplification without droplet motion. This combination of the established real-time reverse transcription-PCR assay with digital microfluidics is ideal for isolating single-copy RNA and virions from a complex environment, and will be useful in viral discovery and gene-profiling applications.

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PDF-file: 21 pages; size: 2 Mbytes

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  • Journal Name: Analytical Chemistry, N/A, no. 6, March 15, 2008, pp. 1854-1858; Journal Issue: 6

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  • Report No.: LLNL-JRNL-400243
  • Grant Number: W-7405-ENG-48
  • Office of Scientific & Technical Information Report Number: 929527
  • Archival Resource Key: ark:/67531/metadc898269

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Office of Scientific & Technical Information Technical Reports

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  • December 19, 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; Wheeler, E; Lee-Houghton, L; Watkins, N; Nasarabadi, S; Hebert, N et al. On-chip single-copy real-time reverse-transcription PCR in isolated picoliter droplets, article, December 19, 2007; Livermore, California. (digital.library.unt.edu/ark:/67531/metadc898269/: accessed August 15, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.