A disposable polydimethylsiloxane (PDMS) microdevices for DNA amplification with low power consumption
A. LekwichaiSupanit PorntheeraphatWin BunjongpruJakrapong SupadechW. SripumkhaiSakon RahongCharndet HruanunAmporn PoyaiJ. Nukeaw
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Polymerase Chain Reaction (PCR) microchip has become an important tool in a molecular biological research. Many researchers have attempt to improve the efficiency of the device in terms of type of materials, fabrication process, temperature control system, design of the device, detection methods, etc. In this study, we demonstrate the disposable polydimethylsiloxane (PDMS) microchip provided for DNA amplification. The devices consists of two mainly parts, first part is PDMS static chamber that casting on Silicon (Si) mold that bonded with thin glass sheet by RF plasma cleaner. The other parts are Pt-microheater and Pt-temperature sensor on SiO 2 /Si substrate. The thin film Pt-microheater and Pt-temperature sensor (RTD) were fabricated with CMOS compatible process. In the results, with 10 ¿l of PCR mixture volume, the approximately 700 bp DNA were successfully amplified within 50 minutes by 32 PCR cycles. The amplified products were comparable with conventional method by agarose gel electrophoresis method. With this PCR chip we can save cost by reducing the reagents volume, time consuming and repeatable fabrication. Further information about microchip fabrication and application will be presented.Keywords:
Microheater
Polydimethylsiloxane
Lab-on-a-Chip
Our precedent study has reported glass-PDMS (polydimethylsiloxane) based biochip for the gene PCR (polymerase chain reaction). To prevent the contamination of bio sample, the once used biochip must not be used repeatedly. However, the fabrication cost of microheater and microsensor of the biochip was not cheap to use it as a disposable chip. This paper proposes new PCR-chip where the glass substrate integrated with the microheater and microsensor is detachable from the reaction chamber where the sample is injected. That makes it possible to reuse the glass substrate repeatedly. The performance of the proposed detachable PCRchip was compared with that of the precedent monolithic PCR-chip. The results showed that the SRY (sex determining Y chromosome) gene PCR was successfully performed in the detachable chip compared with the monolithic chip. However, the more efforts to improve the efficiency of surface treatment of PDMS chip are needed to increase the possibility of applying the detachable chip to the detecting of male infertility.
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Integration of microfluidics technology with DNA microarrays enables building complete sample-to-answer systems that are useful in many applications such as clinic diagnostics. In this chapter, a fully integrated microfluidic device [1] that consists of microfluidic mixers, valves, pumps, channels, chambers, heaters, and a DNA microarray sensor to perform DNA analysis of complex biological sample solutions is present. This device can perform on-chip sample preparation (including magnetic bead-based cell capture, cell preconcentration and purification, and cell lysis) of complex biological sample solutions (such as whole blood), polymerase chain reaction, DNA hybridization, and electrochemical detection. A few novel microfluidic techniques were developed and employed. A micromix-ing technique based on a cavitation microstreaming principle was implemented to enhance target cell capture from whole blood samples using immunomagnetic beads. This technique was also employed to accelerate DNA hybridization reaction. Thermally actuated paraffin-based microvalves were developed to regulate flows. Electrochemical pumps and thermopneumatic pumps were integrated on the chip to provide pumping of liquid solutions. The device is completely self-contained: no external pressure sources, fluid storage, mechanical pumps, or valves are necessary for fluid manipulation, thus eliminating possible sample contamination and simplifying device operation. Pathogenic bacteria detection from ~mL whole blood samples and single-nucleotide polymorphism analysis directly from diluted blood were demonstrated. The device provides a cost-effective solution to direct sample-to-answer genetic analysis, and thus has a potential impact in the fields of point-of-care genetic analysis, environmental testing, and biological warfare agent detection.
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A microheater and a thermal sensor were fabricated inside elastomeric polydimethylsiloxane microchannels by injecting silver paint (or other conductive materials) into the channels. With a high-precision control scheme, microheaters can be used for rapid heating, with precise temperature control and uniform thermal distribution. Using such a microheater and feedback system, a polymerase chain reaction experiment was carried out whereas the DNA was successfully amplified in 25 cycles, with 1 min per cycle.
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Our precedent study has reported glass-PDMS (polydimethylsiloxane) based biochip for the gene PCR (polymerase chain reaction). To prevent the contamination of bio sample, the once used biochip must not be used repeatedly. However, the fabrication cost of microheater and microsensor of the biochip was not cheap to use it as a disposable chip. This paper proposes new PCR-chip where the glass substrate integrated with the microheater and microsensor is detachable from the reaction chamber where the sample is injected. That makes it possible to reuse the glass substrate repeatedly. The performance of the proposed detachable PCRchip was compared with that of the precedent monolithic PCR-chip. The results showed that the SRY (sex determining Y chromosome) gene PCR was successfully performed in the detachable chip compared with the monolithic chip. However, the more efforts to improve the efficiency of surface treatment of PDMS chip are needed to increase the possibility of applying the detachable chip to the detecting of male infertility.
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Lab-on-a-Chip
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