Automated Microdroplet Platform for Sample Manipulation and Polymerase Chain Reaction

We present a fully automated system performing continuous sampling, reagent mixing, and polymerase chain reaction (PCR) in microdroplets transported in immiscible oil. Sample preparation and analysis are totally automated, using an original injection method from a modified 96-well plate layered with...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2006-11, Vol.78 (22), p.7722-7728
Main Authors: Chabert, Max, Dorfman, Kevin D, de Cremoux, Patricia, Roeraade, Johan, Viovy, Jean-Louis
Format: Article
Language:English
Subjects:
Analytical chemistry
Automation
Biological and medical sciences
Biotechnology
Breast cancer
Breast Neoplasms - genetics
Breast Neoplasms - pathology
Cell Line
Cells
Chemistry
chip
device
dna amplification
Exact sciences and technology
Female
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression Profiling - methods
Genetic engineering
Genetic technics
Humans
In vitro gene amplification. Pcr technique
Lasers
Methods. Procedures. Technologies
Microchemistry - methods
multiplex pcr
Oils & fats
Oils - chemistry
Polymerase Chain Reaction
power
Receptors, Progesterone - metabolism
Reproducibility of Results
Spectrometric and optical methods
Spectrometry, Fluorescence - methods
system
Time Factors
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Summary:We present a fully automated system performing continuous sampling, reagent mixing, and polymerase chain reaction (PCR) in microdroplets transported in immiscible oil. Sample preparation and analysis are totally automated, using an original injection method from a modified 96-well plate layered with three superimposed liquid layers and in-capillary laser-induced fluorescence endpoint detection. The process is continuous, allowing sample droplets to be carried uninterruptedly into the reaction zone while new drops are aspirated from the sample plate. Reproducible amplification, negligible cross-contamination, and detection of low sample concentrations were demonstrated on numerous consecutive sample drops. The system, which opens the route to strong reagents and labor savings in high-throughput applications, was validated on the clinically relevant quantification of progesterone receptor gene expression in human breast cancer cell lines.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/ac061205e