以磁珠萃取 DNA 之微流體晶片研製

本研究開發出一種新的微流體DNA萃取，利用玻璃與PCB電路板製作，與其他實驗室所做的晶片迥然不同，將醫院與醫學臨床使用的設備與環境縮小至手掌大小的微流體晶片當中，大幅減少空間。並且利用低成本的PCB電路板取代一般實驗室所用的晶圓，也有效的將製作成本降低，提高其經濟效益達到一次性使用的目的。本磁珠萃取DNA微流體晶片只需搭配不同的試劑就可萃取不同種類的DNA，相較傳統萃取需要1至2天的時間，本微流體晶片只需20至30分鐘就可以萃取出一樣濃度與純度的DNA，並且在試劑上更為安全。利用磁珠的特性可在清洗腔體時，配合強力磁鐵將磁珠固定在腔體底部，避免清洗時帶走DNA。本實驗為了達到減少血液樣本消耗，嘗試了150 uL與200 uL血液之間的差異，與臨床與生技公司發展的DNA萃取機台血液量做比較。大部分人工萃取試劑組與機台自動化萃取的血液量在400 uL至1 mL之間，本實驗室晶片可有效大幅度降低血液用量，不浪費血液與試劑的使用，降低了萃取晶片的成本。DNA純度與濃度的判定藉由吸光值的程度區分，吸光值基本都必須在1.60以上，而實驗結果區間在1.62至2.64之間，代表純度與濃度達到後續可以使用的標準。

關鍵字

磁珠； DNA萃取；實驗室晶片；微流道

並列摘要

This thesis presents a novel microfluidic DNA extraction chip fabricated using printed circuit board (PCB), different from the conventional lab chips. The equipment for DNA extraction used in hospitals and clinics is miniaturized to a chip. For disposable purpose, PCB substrate, replacing the wafer, was used to reduce costs. By the assistance of magnetic beads, different kinds of DNAs can be extracted using different reagents. Under the same concentration and purity DNA, the extraction procedure takes only 20-30 minutes by chip, however, 1-2 days by traditional method. In addition, the reagents needed in this study are much safer. DNAs bound on magnetic beads can be immobilized in the chamber by a permanent magnet during washing, so that DNAs will not be drained away. Four different doses of blood, i.e. 100 uL, 150 uL, 200 uL, and 250 uL, were tried in this study for comparison. The blood consumptions for most manual operation methods or automatic extraction machines are between 400 uL to 1 mL. The necessary amount of blood is significantly reduced using this DNA extraction chip, without wasting blood and reagents. The purity and concentration of the extracted DNA can be determined by the degree of optical absorption, which must be greater than 1.60 basically. Using the microfluidic DNA extraction chip, the OD values reach 1.62 to 2.64.

並列關鍵字

magnetic beads ； DNA extraction ； microchannel ； lab-on-a-chip

參考文獻

[1] Zhang, Wu, and Wang, “Gravitational Sedimentation Induced Blood Delamination for Continuous Plasma Separation on a Microfluidics Chip” Analytical Chemistry,March 20, 2012,p3780-3786

[2] Hosokawa , Ohnishi ,and Fukasawa “A microchip flow-chamber system for quantitative assessment of the platelet thrombus formation process” SciVerse ScienceDirect November ,27, 2011 p154–161

[3] Yang, Forouzan, and Brown ,“Integrated separation of blood plasma from whole blood for microfluidic paper-based analytical devices” Lab on a Chip August ,2011,p 274–280

[4] Shim and Ahn ,“An on-chip whole blood/plasma separator using hetero-packed beads at the inlet of a microchannel”, Lab on a Chip , December ,2011,p863–866

[5] Choi, Song, and Choi, “Continuous blood cell separation by hydrophoretic filtration”, Lab on a Chip, JuLy ,2007, p1532–1538

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以磁珠萃取 DNA 之微流體晶片研製

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