Fabrication of amino silane-coated microchip for DNA extraction from whole blood.
A simple microchip device for DNA extraction was constructed based on electrostatic interactions between surface amine groups and DNA. Microchannel was fabricated on silicon wafer by photolithography and coated with 3-aminopropyltriethoxysilane (APTES) or 3-[2-(2-aminoethylamino)-ethylamino]-propyltrimethoxysilane (AEEA) to introduce amine groups on the surface. Determination of the number of surface amine groups and optimization of DNA capture condition were demonstrated to characterize the microchip. Capacities of capturing DNA were approximately 97 ng/cm2 in APTES and 194 ng/cm2 in AEEA modified microchips, respectively. The amount of DNA captured in the microchip increased depending on surface amine density. Furthermore, DNA extraction using amine-coated microchip from whole blood was examined. Quantification of DNA and proteins in washing or eluting fraction indicates that proteins were removed at washing steps and only DNA was effectively eluted by changing alkalinity of buffer from pH 7.5 to 10.6. The amount of DNA extracted from whole blood was approximately 10 ng and its recovery ratio was 27-40%. Performance of PCR for the eluted fraction indicates that DNA extracted from whole blood was well purified using amine-coated microchip.
Nakagawa
Takahito
T
Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
Tanaka
Tsuyoshi
T
Niwa
Daisuke
D
Osaka
Tetsuya
T
Takeyama
Haruko
H
Matsunaga
Tadashi
T
D000327:Adsorption; D010582:Bacteriophage lambda / Q000235:genetics; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D004279:DNA, Viral / Q000097:blood* / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005029:Ethylenediamines / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D017646:Organosilicon Compounds / Q000737:chemistry*; D011437:Propylamines; D012821:Silanes / Q000737:chemistry*; D055672:Static Electricity; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D004279:DNA, Viral; D005029:Ethylenediamines; D017646:Organosilicon Compounds; D011437:Propylamines; D012821:Silanes; C110457:3-(2-(2-aminoethyl)ethylamino)propyltrimethoxysilane; C477625:amino-propyl-triethoxysilane