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

Journal Article · Wed Dec 19 04:00:00 EST 2007 · Analytical Chemistry, N/A, no. 6, March 15, 2008, pp. 1854-1858

OSTI ID:929527

Beer, N R; Wheeler, E; Lee-Houghton, L; Watkins, N; Nasarabadi, S; Hebert, N; Leung, P; Arnold, D; Bailey, C; Colston, B

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.

• Research Organization: Lawrence Livermore National Laboratory (LLNL), Livermore, CA
• Sponsoring Organization: USDOE
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 929527
• Report Number(s): LLNL-JRNL-400243
• Journal Information: Analytical Chemistry, N/A, no. 6, March 15, 2008, pp. 1854-1858, Journal Name: Analytical Chemistry, N/A, no. 6, March 15, 2008, pp. 1854-1858 Journal Issue: 6
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
AMPLIFICATION
DETECTION
FLUORESCENCE
RNA
SILICA
THERMAL CYCLING
TRANSCRIPTION