Nucleic acid purification using microfabricated silicon structures

Cady, Nathaniel C.; Stelick, Scott; Batt, Carl A.

doi:10.1016/s0956-5663(03)00123-4

Cited by 196 publications

(154 citation statements)

References 16 publications

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“…It is important to note that 200ng DNA overloads the capacity of the microdevice without porous silicon due to the limitation of surface area. Therefore the binding capacity of the microdevice without porous silicon is approximately 75ng/cm 2 which agrees with the results of Cady et al (Cady et al, 2003) who found that the binding capacity of nonporous micropillars was approximate 82ng/cm 2 . The previous researches proved that the performance of DNA extraction microdevice was determined by the surface area of the matrix and the extracted DNA was found to increase linearly with the surface area (Fan et al, 1999;Kwakye et al, 2006).…”

Section: Optimization Of Cell Lysis Processsupporting

confidence: 91%

“…But a high packing density for larger surface area in the microfluidic device results in problems of backpressure and clogging of crude samples, and what is more, it is difficult to control the small particles in microdevices. A micropillar array fabricated by MEMS technology in a microchamber or channel increases the surface area available for DNA adsorption (Christel et al, 1999;Cady et al, 2003). However, the increasing surface area is limited and the problems of clogging could not be completely solved.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

MEMS-Based Microdevice for Cell Lysis and DNA Extraction

Chen¹,

Cui²,

Cai³

et al. 2012

Microelectromechanical Systems and Devices

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Section: Optimization Of Cell Lysis Processsupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

MEMS-Based Microdevice for Cell Lysis and DNA Extraction

Chen¹,

Cui²,

Cai³

et al. 2012

Microelectromechanical Systems and Devices

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“…For example, silica resins have been optimized for DNA capture in microchambers in the presence of chaotropic salts [8,9]. 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].…”

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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“…An example of such useful devices is given by the integrated microfluidic platform, known as the "microFLUIDICS DESKTOP" (Figure 4), developed by Cady and co-workers [185] for detecting Listeria monocytogenes by real time PCR. Monolithic DNA purification/real-time PCR silicon chips ( Figure 5) were fabricated utilizing standard semiconductor processing technologies.…”

Section: Biosensors Microarrays Micro and Nano Electro-mechanical-smentioning

confidence: 99%