非接觸式加熱雙輸出可拋式PCR晶片之研發

Using Non-Contact Heating for the Development of a dual Output Disposable PCR Biochip

林宗漢(Tsung-Han Lin)

指導教授 ： 朱元南

國立臺灣大學/生物資源暨農學院/生物產業機電工程學研究所/碩士(2006年)

https://doi.org/10.6342/NTU.2006.00172

摘要

微流體生物晶片具有反應效率高、試劑用量低、體積小、可大量生產降低成本等優點。基於輻射加熱系統效率較高，槽狀式PCR晶片體積小，本研究結合這兩者的優點，發展非接觸式加熱平行輸出之可拋式PCR生物晶片系統。本研究應用膠膜黏合三片聚酯薄片(PET)，成為槽狀可拋棄式PCR晶片，可以防止蒸發，僅需5.5μl的DNA試劑，即可完成PCR反應，相較於一般商業生產之熱傳導式加熱PCR系統可節省約 ~ PCR試劑用量。本研究採用熱輻射加熱和強制冷卻，升溫速率每秒2.75℃，降溫速率每秒3.5℃，溫度最大過超越量0.65℃，溫度穩態誤差可控制在0.65℃以內，相較於傳統熱傳導式PCR系統之速率快。本研究並實驗二維陣列PCR晶片反應之穩定性，受限於強制熱對流方向是單一方向，導致二維陣列晶片降溫不均勻，經實驗後僅能成功放大一維方向的PCR晶片。

關鍵字

非接觸式加熱PCR ； 可拋式生物晶片

並列摘要

Microfluidic biochips have many advantages including low reagent volume, high reaction efficiency, small size, mass produceability and low cost. The purpose of this study was to develop a non-contact heating, dual output, disposable PCR biochip system, combining the higher thermal efficiency of radiation heating and the advantage of low reagent volume of the well type PCR chips. The study uses a double-sided bonding film to seal three pieces of PET film into a sandwich structure. The inner pieces have pre-manufactured holes to form a closed well with fluid injection and removal ports. This makes a well type disposable PCR biochip. The bonding film could seal up the PCR chip and prevent evaporation. The volume of reagent for a complete PCR reaction was only 5.5μl, only about a half to three fourths that with conventional heating PCR systems. Thanks to the radiation heating and forced convection, the heating rate was 2.75℃/sec ; the cooling rate was 3.5℃/sec, and the thermal transient overshoot and thermal stability were all within 0.65℃/sec. The heating/cooling rate were slightly higher comparing to traditional PCR thermal cyclers .Attempts were made to expand the biochip system to two-dimensional but failed possibly due to uneven temperature distribution among the chips.

並列關鍵字

Non-contact heating PCＲ ； disposable biochip

參考文獻

18. Nakanishi, H., T. Nishimoto, R. Nakamura, A. Yotsumoto.

Chung, D. C. Han and J. K. Chang. 2003. PDMS-based micro

16. Lin, Y. C., M. Y. Huanga, K. C. Young, T. T. Chang and C.

microchambers for DNA amplification. Sens. Actuators A

and J.P. Landers. 2001. Polymerase Chain Reaction in