Fabrications of a continuous-flow DNA amplifier using dry film resist

Kong, Dae Young; Kang, Tae Wook; Seo, Chang Taeg; Cho, Chan Seob; Lee, Jong Hyun

doi:10.1007/s13206-010-4303-9

Cited by 4 publications

(2 citation statements)

References 13 publications

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“…So, the effect on the temperature distribution due to convective heat transfer inside the microchannel is assumed to be negligible when the flow rate of the sample in the microchannel is less than 5 ll/min. Because of the information found in prior literature, 2,5,8,12,14,15,[26][27][28][29][30][31] in order to make sure that the DNA amplification process is successful, the Re of the sample flow inside the microchannel was chosen to be less than 1. In our simulated results, the uniformity of the chip temperature at a specific reaction region is assured when the flow rate of the sample in the microchannel is less than 5 ll/min or the Re of the sample flow is less than 1.…”

Section: Design Conceptmentioning

confidence: 99%

One-heater flow-through polymerase chain reaction device by heat pipes cooling

Chen

Liao

et al. 2015

Biomicrofluidics

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This study describes a novel microfluidic reactor capable of flow-through polymerase chain reactions (PCR). For one-heater PCR devices in previous studies, comprehensive simulations and experiments for the chip geometry and the heater arrangement were usually needed before the fabrication of the device. In order to improve the flexibility of the one-heater PCR device, two heat pipes with one fan are used to create the requisite temperature regions in our device. With the integration of one heater onto the chip, the high temperature required for the denaturation stage can be generated at the chip center. By arranging the heat pipes on the opposite sides of the chip, the low temperature needed for the annealing stage is easy to regulate. Numerical calculations and thermal measurements have shown that the temperature distribution in the five-temperature-region PCR chip would be suitable for DNA amplification. In order to ensure temperature uniformity at specific reaction regions, the Re of the sample flow is less than 1. When the microchannel width increases and then decreases gradually between the denaturation and annealing regions, the extension region located in the enlarged part of the channel can be observed numerically and experimentally. From the simulations, the residence time at the extension region with the enlarged channel is 4.25 times longer than that without an enlarged channel at a flow rate of 2 ll/min. The treated surfaces of the flow-through microchannel are characterized using the water contact angle, while the effects of the hydrophilicity of the treated polydimethylsiloxane (PDMS) microchannels on PCR efficiency are determined using gel electrophoresis. By increasing the hydrophilicity of the channel surface after immersing the PDMS substrates into Tween 20 (20%) or BSA (1 mg/ml) solutions, efficient amplifications of DNA segments were proved to occur in our chip device. To our knowledge, our group is the first to introduce heat pipes into the cooling module that has been designed for a PCR device. The unique architecture utilized in this flow-through PCR device is well applied to a low-cost PCR system. V C 2015 AIP Publishing LLC.

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Section: Design Conceptmentioning

confidence: 99%

One-heater flow-through polymerase chain reaction device by heat pipes cooling

Chen

Liao

et al. 2015

Biomicrofluidics

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“…Recently, dry film photopolymers have moved away from their original purpose of providing sacrificial layers for the fabrication of printed circuit boards and were used for fabricating electroplating moulds [34,35], stamp cavities [36] and replication tools [37,38], for wafer bonding [39,40] and sensor packaging [41], as an etch mask for silicon-DRIE [42] and as a permanent material for microfluidic applications [43][44][45][46]. So far, the most frequently used permanent films are Riston (DuPont) and Ordyl SY (Elga Europe).…”

Section: Selective Sealing Of Silicon Microchannelsmentioning

confidence: 99%

Microfluidics in silicon/polymer technology as a cost-efficient alternative to silicon/glass

Kalkandjiev

Riegger

Kosse

et al. 2011

J. Micromech. Microeng.

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We investigate TMMF photopolymer as a cost-efficient alternative to glass for the leak-tight sealing of high-density silicon microchannels. TMMF enables low temperature sealing and access to structures underneath via lamination and standard UV-lithography instead of costly glass machining and anodic bonding. TMMF is highly transparent and has a low autofluorescence for wavelengths larger than 400 nm. As the photopolymer is too thin for implementing bulky world-to-chip interfaces, we propose adhesive bonding of cyclic olefin copolymer (COC) modules. All materials were tested according ISO 10993-5 and showed no cytotoxic effects on the proliferation of L929 cells. To quantify the cost efficiency of the proposed techniques, we used an established silicon/Pyrex nanoliter dispenser as a reference and replaced structured Pyrex wafers by TMMF laminates and COC modules. Thus, consumable costs, manpower and machine time related to sealing of the microchannels and implementing the world-to-chip interface could be significantly reduced. Leak tightness was proved by applying a pressure of 0.2 MPa for 5 h without delamination or crosstalk between neighboring microchannels located only 100 μm apart. In contrast to anodic bonding, the proposed techniques are tolerant to surface inhomogeneities. They enable manufacturing of silicon/polymer microfluidics at lower costs and without compromising the performance compared to corresponding silicon/glass devices.

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Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens

Zhang

Wang

Xing

2011

Biomed Microdevices

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In the field of continuous-flow PCR, the amplification throughput in a single reaction solution is low and the single-plex PCR is often used. In this work, we reported a flow-based multiplex PCR microfluidic system capable of performing high-throughput and fast DNA amplification for detection of foodborne bacterial pathogens. As a demonstration, the mixture of DNA targets associated with three different foodborne pathogens was included in a single PCR solution. Then, the solution flowed through microchannels incorporated onto three temperature zones in an oscillatory manner. The effect factors of this oscillatory-flow multiplex PCR thermocycling have been demonstrated, including effects of polymerase concentration, cycling times, number of cycles, and DNA template concentration. The experimental results have shown that the oscillatory-flow multiplex PCR, with a volume of only 5 μl, could be completed in about 13 min after 35 cycles (25 cycles) at 100 μl/min (70 μl/min), which is about one-sixth of the time required on the conventional machine (70 min). By using the presently designed DNA sample model, the minimum target concentration that could be detected at 30 μl/min was 9.8 × 10(-2) ng/μl (278-bp, S. enterica), 11.2 × 10(-2) ng/μl (168-bp, E. coli O157: H7), and 2.88 × 10(-2) ng/μl (106-bp, L. monocytogenes), which corresponds to approximately 3.72 × 10(4) copies/μl, 3.58 × 10(4) copies/μl, and 1.79 × 10(4) copies/μl, respectively. This level of speed and sensitivity is comparable to that achievable in most other continuous-flow PCR systems. In addition, the four individual channels were used to achieve multi-target PCR analysis of three different DNA samples from different food sources in parallel, thereby achieving another level of multiplexing.

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Fabrications of a continuous-flow DNA amplifier using dry film resist

Cited by 4 publications

References 13 publications

One-heater flow-through polymerase chain reaction device by heat pipes cooling

One-heater flow-through polymerase chain reaction device by heat pipes cooling

Microfluidics in silicon/polymer technology as a cost-efficient alternative to silicon/glass

Multichannel oscillatory-flow multiplex PCR microfluidics for high-throughput and fast detection of foodborne bacterial pathogens

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