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Anal Chem. 2008 Mar 15;80(6):1854-8. doi: 10.1021/ac800048k. Epub 2008 Feb 16.

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

Author information

1
Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, California 94551, USA. beer2@llnl.gov

Abstract

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.

PMID:
18278951
DOI:
10.1021/ac800048k
[Indexed for MEDLINE]

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