title: Solid phase DNA extraction on PDMS and direct amplification.
abstract: In this paper we report an innovative use of Poly(DiMethyl)Siloxane (PDMS) to design a microfluidic device that combines, for the first time, in one single reaction chamber, DNA purification from a complex biological sample (blood) without elution and PCR without surface passivation agents. This result is achieved by exploiting the spontaneous chemical structure and nanomorphology of the material after casting. The observed surface organization leads to spontaneous DNA adsorption. This property allows on-chip complete protocols of purification of complex biological samples to be performed directly, starting from cells lysis. Amplification by PCR is performed directly on the adsorbed DNA, avoiding the elution process that is normally required by DNA purification protocols. The use of one single microfluidic volume for both DNA purification and amplification dramatically simplifies the structure of microfluidic devices for DNA preparation. X-Ray Photoelectron Spectroscopy (XPS) was used to analyze the surface chemical composition. Atomic Force Microscopy (AFM) and Field Emission Scanning Electron Microscopy (FESEM) were employed to assess the morphological nanostructure of the PDMS-chips. A confocal fluorescence analysis was utilized to check DNA distribution inside the chip.
authors: FBK-Fondazione Bruno Kessler, Center for Materials and Microsystems, Povo, Trento, Italy. pasqua@fbk.eu
authors: Laura
authors: L
authors: Pasquardini
authors: Cristina
authors: C
authors: Potrich
authors: Marzia
authors: M
authors: Quaglio
authors: Andrea
authors: A
authors: Lamberti
authors: Salvatore
authors: S
authors: Guastella
authors: Lorenzo
authors: L
authors: Lunelli
authors: Matteo
authors: M
authors: Cocuzza
authors: Lia
authors: L
authors: Vanzetti
authors: Candido Fabrizio
authors: CF
authors: Pirri
authors: Cecilia
authors: C
authors: Pederzolli
mesh_terms: D004247:DNA / Q000097:blood* / Q000302:isolation & purification; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D018625:Microscopy, Atomic Force; D056951:Photoelectron Spectroscopy
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon; D004247:DNA