聚合酶連鎖反應晶片之研究及其製程分析

Abstract

聚合酶連鎖反應(Polymerase Chain Reaction, PCR)於1986年發表至今，使得基因工程研究的發展迅速。再加上近年來微機電技術(Micro-Electro-Mechanical System, MEMS)蓬勃發展下，使得聚合酶連鎖反應可藉由微機電技術，達到縮小尺寸，減低能源消耗，卻能加快聚合酶連鎖反應反應的進行。 由於聚合酶連鎖反應需要固定的溫度來進行，為了使傳熱快速，使用矽晶圓作為聚合酶連鎖反應晶片的開發材料，並可以利用微機電製程中的黃光微影製程、乾蝕刻製程及陽極接合製程，來製作縮小化的聚合酶連鎖反應晶片，為本研究之主要目的。另外，本研究亦利用模擬分析進行晶片的熱分佈分析，並改進因聚合酶連鎖反應所需之三個不同溫度區之溫度干擾問題。 首先本研究利用微機電技術及聚二甲基矽氧烷製程嘗試製作第一代微流道晶片，並由測試結果中可以得知第一代微流道晶片可以簡化溫控裝置，但仍有DNA反應物滲漏之情形。所以第二代微流道晶片則利用微機電技術把加熱電極及溫度感測電極製作於晶片上，並利用陽極接合封裝晶片，解決反應物滲漏之問題。最後本研究利用模擬分析軟體求出絕熱區之最佳化寬度及深度，用於減少時間成本下亦能夠得到相同的絕熱效果。 由測試結果可以得知，聚合酶連鎖反應晶片內的DNA反應物確實能夠被加熱至所需之溫度，並進行聚合酶連鎖反應。但因微流道注入孔無法和外界注射器緊密結合，造成反應物注入時的浪費，希望未來能由晶片與外界器材之封裝來改進整個晶片。

Polymerase Chain Reaction (PCR) was issued so far in 1986; it made genetic engineering technology fast growing. Because Micro-Electro-Mechanical System (MEMS) grows vigorously in recent years, it has made Polymerase Chain Reaction change significantly in reducing size scale, lowering energy consumption, and speeding up the process. The process of Polymerase Chain Reaction needed fixed and stable temperature. In order to make heat transfer faster, using the silicon wafer as the material of PCR chip can do the following: utilizing photolithography process, etching process and anodic bonding process in MEMS process, to reduce the size of PCR chip. This is the main purpose of this research. This research also utilizes simulation to analyze the heat distribution of the chip, and to solve the interference question of temperature between three different temperature zones of the PCR chip. The research begins with using MEMS technology and PDMS process attempting to make the first micro-flow chip. The results had shown it could simplify the temperature controlling device, but sometimes the DNA reactant would seep from the joint of silicon and PDMS. Thus the heaters and temperature sensors were plated with Pt and Cr to the second edition micro-flow chip by MEMS technology and packaged the pyrex 7740 glass and silicon wafer by anodic bonding process to solve the problem of reactant seepage. Finally, simulation software has found the optimal width and depth of the isolation zone. From the testing results, DNA reactant in PCR Chip can really be heated to appropriate temperature, and be in process of Polymerase Chain Reaction. Because the injection hole at the PCR chip was unable to connect closely with external syringe, it causes reactant waste when inject into the PCR chip. Research should continue on the encapsulation of PCR chip in order to solve the problem mentioned above.