Solid phase DNA extraction on PDMS and direct amplification

被引:36
|
作者
Pasquardini, Laura [1 ]
Potrich, Cristina [1 ,2 ]
Quaglio, Marzia [3 ]
Lamberti, Andrea [3 ,5 ]
Guastella, Salvatore [5 ]
Lunelli, Lorenzo [1 ,2 ]
Cocuzza, Matteo [4 ,5 ]
Vanzetti, Lia [1 ]
Pirri, Candido Fabrizio [3 ,5 ]
Pederzolli, Cecilia [1 ]
机构
[1] FBK Fdn Bruno Kessler, Ctr Mat & Microsyst, I-38123 Povo, Trento, Italy
[2] CNR Consiglio Nazl Ric, Ist Biofis, I-38123 Povo, Trento, Italy
[3] CSHR Ctr Human Space Robot, IIT Fdn Ist Italiano Tecnol, I-10129 Turin, Italy
[4] CNR IMEM, I-43124 Parma, Italy
[5] Politecn Torino, Mat Sci Chem Engn DISMIC Dept, Chilab Mat & Microsyst Lab, I-10129 Turin, Italy
关键词
REAL-TIME PCR; SURFACE MODIFICATION; POLY(DIMETHYLSILOXANE); CHIP; FABRICATION; MICROCHIP; SYSTEMS;
D O I
10.1039/c1lc20371a
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:4029 / 4035
页数:7
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