Fabrication of amino silane-coated microchip for DNA extraction from whole blood

Nakagawa, Takahito; Tanaka, Tsuyoshi; Niwa, Daisuke; Osaka, Tetsuya; Takeyama, Haruko; Matsunaga, Tadashi

doi:10.1016/j.jbiotec.2004.08.018

Cited by 123 publications

(118 citation statements)

References 24 publications

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“…Another approach has been to use microfabricated silica structures etched into a surface to increase surface area [10]. DNA binding molecules such as amino silanes have been coated onto glass slides to improve capacity in extraction devices and can release the DNA at high pH [11]. Other polyamine coatings such as polylysine have been used to coat microscope slides for making DNA microarrays, but these are not designed to release the DNA.…”

Section: Discussionmentioning

confidence: 99%

Capture of genomic DNA on glass microscope slides

Nanassy

Haydock

Reed

2007

Analytical Biochemistry

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It is well known that DNA strands bind to silica surfaces in the presence of high concentrations of chaotropic salts. We developed simple methods to evaluate binding and recovery of DNA on flat glass microscope slides and compared their properties to commercially available silica "spin columns". Surprisingly, genomic DNA was recovered efficiently from untreated glass slides. Binding and elution times from glass slides were optimized in experiments with DNA samples of various sizes and defined buffers. Average DNA recovery from 500 ng of input genomic DNA varied from 25-53 % for the glass slide protocol. Yields were comparable to spin columns. Human serum albumin and plasma components decreased DNA binding to glass several-fold in a concentration dependent manner. These results support the concept of using flat glass slides as DNA purification surfaces in microfluidics devices for PCR sample preparation. Keywords DNA extraction; silica surfaces; PCR; microfluidicsThe use of powdered glass or other high surface area silica materials has been widely adopted as a basic technique for purifying DNA from biological samples [1]. DNA strands bind efficiently to silica particles in the presence of high concentrations of chaotropic salts such as guanidinium thiocyanate or sodium perchlorate. After washing away contaminating substances with high salt buffers, the bound DNA is released from the silica surface in the presence of water or low salt buffer. The recovered DNA is suitable for use in PCR or other amplification methods. These methods are used extensively in medical diagnostics and forensics, in recombinant technology and for molecular genetics.Silica-based DNA purification is compatible with microfluidic diagnostic devices. Such "lab on a chip" devices may help bring the benefits of PCR-based molecular diagnosis to users and point-of-care settings that lack the capacity for standard laboratory-based DNA purification [2] [3]. In such applications there are significant pressures to keep costs low. Therefore, DNA purification devices must be designed to use the least expensive materials possible, and without unnecessary engineering. Early researchers showed that the curved inner surfaces of glass test tubes could be used to purify DNA from agarose gels, but their binding capacity was deemed too small for most preparative experiments [4]. Currently, it is standard practice to maximize DNA binding surface area, for example by using silica glass bead matrices. However, such engineering measures may not be necessary to achieve satisfactory DNA purification yields.

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Section: Discussionmentioning

confidence: 99%

Capture of genomic DNA on glass microscope slides

Nanassy

Haydock

Reed

2007

Analytical Biochemistry

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“…For efficient recovery of the photocleaved cDNA with a low background, an APTESmodified glass was used as an adsorption substrate of the cDNA [16]. Prior to the photorecovery experiment, adsorption of DNAs onto the APTES substrate was investigated in a series of buffers with different pHs.…”

Section: Preparation Of Aptes-modified Glass and Examination Of Cdna mentioning

confidence: 99%

Photoassisted Recovery of DNA Molecules for On-chip Directed Evolution

Ueno

Ono

Tanaka

et al. 2012

J. Photopol. Sci. Technol.

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Directed evolution is a powerful approach to creating functional biomolecules, and a combinative use of microarray chip technology is expected to have a great potential for realizing a sophisticated directed evolution with higher speed and quantitative performance than ever before. Although "recovery" of evaluated and selected molecules followed by amplification is an essential step in the performance of directed evolution, conventional DNA or protein microarray chips are limited only to analytical use. In this study, we have developed a novel technology for DNA microarray that is applicable to an artificial Darwinian selection on a chip. DNA molecules were microarrayed on a gold surface via an oligonucleotide linker that contains a nitrobenzyl group as a photocleavable moiety and were recovered from the surface by spot-selective photoirradiation at few micrometer precision. The recovered DNA molecules were confirmed by polymerase chain reaction. Thus, in this study, we have realized a new function of biomolecular microarray technology that is useful for the establishment of on-chip directed molecular evolution.

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“…16,17 Recently, monolayers of functionalized silanes and epoxysilanes have attracted much attention for anchoring biomolecules such as proteins, enzymes and DNAs. [18][19][20] These silanes can act as cross-linkers because they contain Si-O bonds that react with surface hydroxyl groups of metal oxides, glass, silicon, quartz etc. and pendant-functionalized hydrocarbon chains that can bind with organic materials and desired biomolecules.…”

Section: -8mentioning

confidence: 99%

Biocompatible self-assembled monolayer platform based on (3-glycidoxypropyl)trimethoxysilane for total cholesterol estimation

Kumar¹,

Singh²,

Agrawal³

et al. 2011

Anal. Methods

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An electrochemical cholesterol biosensor has been fabricated via covalent immobilization of cholesterol oxidase and cholesterol esterase (ChOx and ChEt) onto a biocompatible self assembled monolayer (SAM) of (3-glycidoxypropyl)trimethoxysilane (GPTMS) fabricated onto an indium tin oxide (ITO) electrode. The ChOx/GPTMS/ITO bioelectrode has been characterized using Fourier transform infrared spectroscopy (FTIR), contact angle measurements (CA), atomic force microscopy (AFM), electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV) techniques. The results of the electrochemical response studies indicate that the ChOx-ChEt/GPTMS/ITO biosensing electrode can be used to estimate the total cholesterol from 1.5 to 6.1 mM, has sensitivity of 0.351 mA (mg/dl) À1, and can be used more than 10 times with a shelf life of up to 10 weeks. The value of the MichaelisMenten constant (K m ) observed as 0.43 mM reveals a strong binding to the biocompatible GPTMS organic platform and shows low interference effect. Further, this biosensor can be used to estimate the total cholesterol concentration in serum samples.

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Fabrication of amino silane-coated microchip for DNA extraction from whole blood

Cited by 123 publications

References 24 publications

Capture of genomic DNA on glass microscope slides

Capture of genomic DNA on glass microscope slides

Photoassisted Recovery of DNA Molecules for On-chip Directed Evolution

Biocompatible self-assembled monolayer platform based on (3-glycidoxypropyl)trimethoxysilane for total cholesterol estimation

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