On-chip single-copy real-time reverse-transcription PCR in isolated picoliter droplets.
Anal Chem
; 80(6): 1854-8, 2008 Mar 15.
Article
em En
| MEDLINE
| ID: mdl-18278951
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article