Thermoplastic Microfluidic Device for On-Chip Purification of Nucleic Acids for Disposable Diagnostics

Bhattacharyya, Arpita; Klapperich, Catherine M.

doi:10.1021/ac051449j

Cited by 145 publications

(142 citation statements)

References 16 publications

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“…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. Porous polymers with embedded silica particles have also been reported [12].…”

Section: Discussionmentioning

confidence: 98%

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: 98%

Capture of genomic DNA on glass microscope slides

Nanassy

Haydock

Reed

2007

Analytical Biochemistry

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“…30,31 Similar to Breadmore's approach, Bhattacharyya et al used a porous reversed-phase monolith in a polymeric device to immobilize the silica beads. 32 However, in this system, the monolith made no contribution to the DNA extraction, and the limited accessibility of silica particles yielded an extraction efficiency of ∼70% for the first extraction with a loading capacity of <40 ng λ-phage DNA. Although extraction efficiency decreased rapidly after the first run, primarily because of the unstable structure of the monolith and beads, this concern becomes irrelevant in singleuse, disposable devices.…”

Section: Organic Polymeric Monolithsmentioning

confidence: 95%

Purification of Nucleic Acids in Microfluidic Devices

et al. 2008

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“…To validate the capability of fabricated channels as microfluidic devices, a "Y" design was used for fluid flow and theoretically compared with the literature (Figures 3(a)-3(c)). 5,8,10,11,14,16,18,20,[37][38][39][40][41][42][43] Other geometries were also fabricated (Figures 3(d) and 3(e)) to show the versatility of the targeted asymmetric calendaring and hydrophobization (TACH) method. The microfluidic devices were manufactured using paper and tested with dyed water (food color) for visualization.…”

Section: B Channel Fabricationmentioning

confidence: 99%

Paper-based microfluidic devices by asymmetric calendaring

et al. 2017

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We report a simple, efficient, one-step, affordable method to produce open-channel paper-based microfluidic channels. One surface of a sheet of paper is selectively calendared, with concomitant hydrophobization, to create the microfluidic channel. Our method involves asymmetric mechanical modification of a paper surface using a rolling ball (ball-point pen) under a controlled amount of applied stress (r z ) to ascertain that only one side is modified. A lubricating solvent (hexane) aids in the selective deformation. The lubricant also serves as a carrier for a perfluoroalkyl trichlorosilane allowing the channel to be made hydrophobic as it is formed. For brevity and clarity, we abbreviated this method as TACH (Targeted Asymmetric Calendaring and Hydrophobization). We demonstrate that TACH can be used to reliably produce channels of variable widths (size of the ball) and depths (number of passes), without affecting the nonworking surface of the paper. Using tomography, we demonstrate that these channels can vary from 10s to 100s of microns in diameter. The created hydrophobic barrier extends around the channel through wicking to ensure no leakages. We demonstrate, through modeling and fabrication, that flow properties of the resulting channels are analogous to conventional devices and are tunable based on associated dimensionless numbers. Published by AIP Publishing. [http://dx

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Thermoplastic Microfluidic Device for On-Chip Purification of Nucleic Acids for Disposable Diagnostics

Cited by 145 publications

References 16 publications

Capture of genomic DNA on glass microscope slides

Capture of genomic DNA on glass microscope slides

Purification of Nucleic Acids in Microfluidic Devices

Paper-based microfluidic devices by asymmetric calendaring

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