Automated microdroplet platform for sample manipulation and polymerase chain reaction

被引:88
作者
Chabert, Max
Dorfman, Kevin D.
de Cremoux, Patricia
Roeraade, Johan
Viovy, Jean-Louis
机构
[1] Inst Curie, Sect Rech, CNRS, UMR 158,IC,Lab Physicochim Curie, F-75005 Paris, France
[2] Inst Curie, Unite Pharmacol, F-75005 Paris, France
[3] Royal Inst Technol, Dept Analyt Chem, SE-10044 Stockholm, Sweden
关键词
D O I
10.1021/ac061205e
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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.
引用
收藏
页码:7722 / 7728
页数:7
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