Abstract
Polymerase chain reaction (PCR) is a technique for nucleic acid amplification, which has been widely used in molecular biology. Owing to the limitations such as large size, high power consumption, and complicated operation, PCR is only used in hospitals or research institutions. To meet the requirements of portable applications, we developed a fast, battery-powered, portable device for PCR amplification and end-point detection. The device consisted of a PCR thermal control system, PCR reaction chip, and fluorescence detection system. The PCR thermal control system was formed by a thermal control chip and external drive circuits. Thin-film heaters and resistance temperature detectors (RTDs) were fabricated on the thermal control chip and were regulated with external drive circuits. The average heating rate was 32 °C/s and the average cooling rate was 7.5 °C/s. The disposable reaction chips were fabricated using a silicon substrate, silicone rubber, and quartz plate. The fluorescence detection system consisted a complementary metal-oxide-semiconductor (CMOS) camera, an LED, and mirror units. The device was driven by a 24 V Li-ion battery. We amplified HPV16E6 genomic DNA using our device and achieved satisfactory results.
Highlights
Polymerase chain reaction (PCR) is a powerful technique to amplify the specific DNA fragments in vitro, which can produce a large amount of target DNA from a very small amount of template DNA in 2 h [1,2,3]
To evaluate the performance of the thin-film heater and the resistance temperature detectors (RTDs), we carried out several experiments to verify the thermal uniformity, accuracy, and precision of the temperature measurement
We used fluorescein to verify the reliability of the reaction chip and fluorescence detection system; the fluorescein reagent was injected into the top-right chamber of the PCR reaction detection thewater fluorescein reagent was injected into the top-right chamber of the reaction chip and system; deionized was injected into the bottom-left chamber using a pipette
Summary
Polymerase chain reaction (PCR) is a powerful technique to amplify the specific DNA fragments in vitro, which can produce a large amount of target DNA from a very small amount of template DNA in 2 h [1,2,3]. It is necessary to develop a portable PCR device for outdoor clinical diagnoses, which has a small size and features battery-powered operation and user-friendly control. Peltier heaters are commonly used as the heating component [12,13,14] They usually need a high-power supply and exhibit poor thermal dissipation, thereby requiring an external fan for cooling. Jeong et al developed a portable low-power thermal cycler based on Pt thin-film heater to amplify Escherichia coli genomic DNA and the PCR results were detected using gel electrophoresis. Both of these methods lacked a miniaturized detection system, which blocks their utilization in portable applications [25]. We used HPV16E6 genomic DNA for the PCR experiment under appropriate conditions and analyzed the results after the last cycle
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