單股DNA於微奈米溝槽電極之基因晶片上固定化與雜交效率之最佳化; Optimization of Immobilization and Hybridization Efficiency of Single-Strand DNA on Micro-Electrode Biochips

NCUIR > Engineering College > National Central University Department of Chemical & Materials Engineering > Electronic Thesis & Dissertation > Item 987654321/3897

Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/3897

Title:	單股DNA於微奈米溝槽電極之基因晶片上固定化與雜交效率之最佳化;Optimization of Immobilization and Hybridization Efficiency of Single-Strand DNA on Micro-Electrode Biochips
Authors:	王志豪;Chih-Hao Wang
Contributors:	化學工程與材料工程研究所
Keywords:	金奈米;去氧核醣核酸;電性偵測;基因晶片;Gold nanoparticles;DNA;Electrical detection;Gene chip
Date:	2005-06-02
Issue Date:	2009-09-21 12:25:25 (UTC+8)
Publisher:	國立中央大學圖書館
Abstract:	本研究主要是利用表面改質與生物分子固定化來建立一個去氧核醣核酸分子檢測界面，並結合半導體製程及電路設計與量測，來研究一簡單新型的電子式基因晶片。整個研究的主題分為兩個部分，第一部分是探討去氧核醣核酸分子的固定化之驗證與最佳化，第二部分是探討以電性量測於去氧核醣核酸分子有雜交與無雜交的電性量測差異性之提升。藉由接觸角量測(contact angle measurement)、原子力顯微鏡(AFM)和表面化學元素分析儀(ESCA)來驗證去氧核醣核酸分子固定化，並利用HP4284量測不同條件下，有雜交與無雜交於微奈米溝槽內阻抗值之差異性。在去氧核醣核酸分子固定化中，我們將二氧化矽表面經過適當的化學改質修飾後，固定去氧核醣核酸分子於表面上。實驗結果顯示，每一步化學改質與固定化反應均得驗證，並於pH為6.6，鹽濃度為0.3M的磷酸緩衝溶液中，固定化與雜交效率可達九成，且固定化與雜交反應時間僅需6小時與3小時即可達反應平衡。在電性測量測有無雜交於微奈米溝槽之差異性，實驗中選擇不同金奈米粒徑之標定、不同寬度之溝槽與量測頻率進行檢測。由電性量測訊號(阻抗值)可發現於頻率之影響較不明顯，但於金奈米粒徑為20nm與溝槽寬度為5 The research aimed at the optimizing the conditions for the immobilization of single-stranded DNA, hybridization of complementary ssDNA and detection of hybridization on the chip by electrical impedance. We used contact angle measurement, atomic force microscopy (AFM), electron spectroscopy for chemical analysis (ESCA) and UV-VIS to verify ssDNA immobilization and hybridization efficiency. Furthermore, gold nanoparticles were used to enhance the signal of the electrical impedance. In this study, we selected different frequency of the impedance measurement, different width of the gap and the sizes of the nanoparticles to reveal the optimum operation condition of the electrical gene chip. As a result, at the frequency of 100k Hz, particle size of 20nm and the gap width of 5mm, the chip has the difference of 16 times by electrical impedance in hybridization and non-hybridization.
Appears in Collections:	[National Central University Department of Chemical & Materials Engineering] Electronic Thesis & Dissertation

Files in This Item:

File	Size	Format

社群 sharing

Loading...