研究目的
To propose and evaluate an optimal structure for a PCB-based micro PCR chip using black PCB and double-sided tapes to minimize noise in fluorescence detection and enable real-time PCR monitoring.
研究成果
The use of a black PCB with white silk printing significantly reduces noise in fluorescence detection, enabling effective real-time PCR monitoring. Among the tested double-sided tapes, three successfully completed PCR, with performance variations observed. The Bhattacharyya distance provided a better exponential increase curve for fluorescence, similar to standard real-time PCR, suggesting the chip structure is viable for quantitative analysis.
研究不足
The study is limited by the specific types of double-sided tapes used, which may not represent all available options. The fluorescence detection method relies on external imaging equipment, which could introduce variability. The chip design may have thermal and mechanical constraints under high-temperature PCR conditions.
1：Experimental Design and Method Selection:
The study designed a micro-PCR chip with a four-layer structure using black PCB and various double-sided tapes to reduce light reflection and noise. Fluorescence detection was performed using a blue LED and DSLR camera for real-time monitoring.
2：Sample Selection and Data Sources:
DNA from Chlamydia trachomatis was used as the sample, with specific primers and probes. Six types of reaction chambers made from different double-sided tapes were tested.
3：List of Experimental Equipment and Materials:
Equipment includes a PIC18F4550 microcontroller, heater, fan, thermistor, blue LED (9600 mcd), DSLR camera (Canon 1100D), and various tapes (e.g., from 3M, Bridge Chemical, Core Tech, Nitto Denko). Materials include black PCB, double-sided tapes with acrylic adhesive, PP cover film, and PCR reagents.
4：Experimental Procedures and Operational Workflow:
The PCR protocol involved 40 cycles of denaturation, renaturation, and extension. Fluorescence images were captured 10 seconds before the last extension step in each cycle, with LED illumination controlled to prevent quenching. Experiments were conducted in quadruplicate for each tape type.
5：Data Analysis Methods:
Fluorescence brightness was analyzed using mean values and Bhattacharyya distance for histogram similarity to compare PCR performance across different tapes.
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