Identification of PCR Product by double-strand DNA Denaturation using Label-free Field-effect Transistor

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We demonstrate an innovative method of PCR product identification that is based on thermal denaturation of PCR products which are immobilized on microbeads using integrated BioFET. Purification step was finished simply by capturing the biotinylated PCR products on 2.8 µm streptavidin magnetic beads and washing with buffer. The suspension of PCR product captured beads was injected to BioFET sensor chamber. After raising temperature upto denaturation point, surface potential of BioFET is changed by mobile DNA in solution from PCR product on microbeads. By differentiating potential change with respect to temperature change, melting temperature (Tm) can be determined. Thermal denaturation of 412bp PCR product results in increase of surface potential and the maximum peak of the differentiated surface potential, so-called Tm is calculated (Tm =47.6 ). We also discriminated three kinds of PCR product by melting temperature analysis (Tm=44 , 47.6 and 53 each). Using integrated BioFET and bead-based easy sample preparation, we could, for the first time to the best of our knowledge, demonstrate identification method of PCR product by electrical label-free detection. As such, our approach establishes a foundation for PCR based biotechnologies with microelectronics.

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Journal: TechConnect Briefs
Volume: 2, Nanotechnology 2008: Life Sciences, Medicine & Bio Materials – Technical Proceedings of the 2008 NSTI Nanotechnology Conference and Trade Show, Volume 2
Published: June 1, 2008
Pages: 632 - 635
Industry sectors: Medical & Biotech | Sensors, MEMS, Electronics
Topic: Chemical, Physical & Bio-Sensors
ISBN: 978-1-4200-8504-4