本文實驗利用製備具有功能性磁性奈米材料,以及分離純化所使用分解溶液、洗滌溶液以及析出溶液,利用磁性材料與溶液的搭配使用,發展出一套高效率自動化核酸純化試劑,應用於人類血液、組織以及細胞中,從中萃取純化DNA與RNA。首先我們先合出具有超順磁性的水相四氧化三鐵,並且利用十八烯酸(Oleic Acid)修飾,讓奈米粒子能夠均勻分散在溶液中,提升純化效率;接著再製備純化試劑,首先配置具有分解細胞效果的分解溶液,讓DNA與RNA可以裸露出來,由於磁性奈米粒子的表面是帶有負電荷,因此利用分解溶液中的Guanidine thiocyante 作為架橋,幫助DNA和RNA與磁珠結合,之後配置洗滌溶液,目的是為了將吸附在磁珠上的蛋白質及不純物質,藉由洗滌的步驟,將不純物質去除,最後利用析出溶液的配製,以pH值調控的方式將吸附於磁珠上的DNA或RNA脫附下來。這套自動化的設備,在施以外加磁場的情況下,不需要離心的動作即可在數十分鐘內完成多組血液樣品、組織以及細胞中DNA與RNA的純化過程,除了能夠在醫療的診斷上節省時間,也能夠節省人力資源,最後希望這套自動化純化核酸試劑,廣泛應用於疾病的檢測、親子鑑定、標靶治療上。
In this work, we described a simple and convenient process 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 capcity of DNA and RNA. Second, The nanoparticles were characterized by transmission electron microscopy, fourier transformation infrared spectroscopy, X-ray diffraction, supercunducting 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. Finally, the results indicate that the magnetic separation can be applied for clinical diagnoses.