學研合作計畫---功能化奈米粒子結合質譜儀運用在檢驗與診斷技術平台的開發(II)
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Date
2010-07-31
Authors
陳家俊
蘇五洲
陳玉如
張瑛芝
陳進勛
林俊成
吳登強
巫坤品
許聞廉
Journal Title
Journal ISSN
Volume Title
Publisher
行政院國家科學委員會
Abstract
在二十一世紀，人類的基因體資料被廣泛的應用在診斷疾病及其他用途上。本計畫的目標，就是發展一系列的技術，希望以後可以使用在偵測人類基因體。在本研究中，我們發展出利用修飾過油酸的磁珠，可以簡單和快速的分離純化DNA與RNA。首先，我們先製備具有超順磁性的水相四氧化三鐵，並且利用油酸修飾，讓磁性奈米粒子能夠均勻分散在溶液中，而表面更具有電荷，以提升純化效率。製備後的磁性奈米粒子，使用電子顯微鏡、傅立葉轉換紅外光譜、X光繞射儀和超導量子干涉磁量儀來鑑定材料的特性。我們也製備各種不同純化試劑，如:血液DNA自動純化試劑、組織DNA自動純化試劑、血液RNA自動純化試劑和細胞RNA自動純化試劑。這些試劑包含了分解液、清洗液及脫附液。血液DNA自動純化試劑和組織DNA自動純化試劑可被用來純化人體血液和組織中的DNA。血液RNA自動純化試劑和細胞RNA自動純化試劑可用於純化人體血液及細胞的RNA。更進一步的，我們用膠體電泳、紫外-可見光儀和聚合&；#37238；鏈鎖反應分析純化後的DNA和RNA。結果顯示，這套自動化純化的方法可以被用來純化DNA和RNA。
In the 21st century, the human genome data has been widely applied to acquire new disease marker and other new contents. The goal of this project is to develop a series of the technology for future genomic diagnostics. In this work, we described a simple and convenient method for the separation of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) with functionalized oleic acid-coated magnetic nanoparticles. First, the magnetic particles of iron oxide (Fe3O4) were coated with oleic acid to enhance the adsorptive capacity of DNA and RNA. Second, the magnetic nanoparticles were characterized by transmission electron microscopy, Fourier transformation infrared spectroscopy, X-ray diffraction, superconducting quantum interference device. We also prepared solution kits such as blood DNA auto kit, tissue DNA auto kit, blood RNA auto kit and cell RNA auto kit. The kits include lysis buffer, wash buffer and elution buffer. The blood DNA auto kit and the tissue DNA auto kit have been used in the separation of DNA from human blood and human tissue, respectively. The blood RNA auto kit and the cell RNA auto kit have been individually used in the separation of RNA from human blood and mammalian cells. Furthermore, we analyzed the purified productions of DNA and RNA with gel electrophoresis, micro-volume UV-vis spectrophotometer and polymerase chain reaction. The results suggest that the method can be used for the purification of DNA and RNA.
In the 21st century, the human genome data has been widely applied to acquire new disease marker and other new contents. The goal of this project is to develop a series of the technology for future genomic diagnostics. In this work, we described a simple and convenient method for the separation of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) with functionalized oleic acid-coated magnetic nanoparticles. First, the magnetic particles of iron oxide (Fe3O4) were coated with oleic acid to enhance the adsorptive capacity of DNA and RNA. Second, the magnetic nanoparticles were characterized by transmission electron microscopy, Fourier transformation infrared spectroscopy, X-ray diffraction, superconducting quantum interference device. We also prepared solution kits such as blood DNA auto kit, tissue DNA auto kit, blood RNA auto kit and cell RNA auto kit. The kits include lysis buffer, wash buffer and elution buffer. The blood DNA auto kit and the tissue DNA auto kit have been used in the separation of DNA from human blood and human tissue, respectively. The blood RNA auto kit and the cell RNA auto kit have been individually used in the separation of RNA from human blood and mammalian cells. Furthermore, we analyzed the purified productions of DNA and RNA with gel electrophoresis, micro-volume UV-vis spectrophotometer and polymerase chain reaction. The results suggest that the method can be used for the purification of DNA and RNA.