Synthesis and surface modification of graphite oxide-cellulose composites for solid phase DNA extraction applications
Başlık çevirisi mevcut değil.
- Tez No: 403438
- Danışmanlar: Prof. NAGAHIRO SAITO
- Tez Türü: Doktora
- Konular: Mühendislik Bilimleri, Engineering Sciences
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2015
- Dil: İngilizce
- Üniversite: Nagoya University
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: 105
Özet
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Özet (Çeviri)
The extraction of DNA is the most vital technique which is widely used in genetic engineering. It is the key process of the recovery, purification and industrial applications of DNA. Generally, silica matrices are the most utilized solid support material due to their unique property of selective DNA binding. The major advantage of silica matrices belongs to its capability to fix DNA on a solid support in a quick and convenient process. However, the adsorption force of DNA based Si-OH functional groups is much greater than that of desorption force, as a result, the DNA extraction efficiency of silica surfaces is limited. Chapter 1 began with a general introduction of various DNA extraction methods, protocol types, fundamentals and applications. The purpose of this study is to invent a new material with suitable functional groups for high DNA extraction efficiency. Therefore, in this study, I have proposed Graphite oxide (GO)/Cellulose composite as an innovative solid based support for DNA extraction. Graphite oxide production and DNA characterization methods and devices are explained in Chapter 2. In chapter 3, my study focused on preparing GO/Cellulose composite and applying single strand DNA (ssDNA) on the surface. The concentration of GO within the composites were ranged from 0 – 6 wt. %. The highest binding capacity was achieved with 4.15 wt. % GO, where the extraction efficiency was reported as 660.4 ng/μl. The results were compared to commercial silica spin column and showed that the extraction efficiency was 50% higher than that of conventional silica column with similar DNA purity. In order to examine the extraction efficiency of DNA in practical usage, genomic DNA samples were applied. In Chapter 4, the extraction of genomic DNA by GO/Cellulose composite was reported. Genomic DNA samples were obtained from forensic DNA sources such as cigarette bud paper, nail, chewing gum, animal tissue and human hair. The bulk % of Graphite oxide was fixed at 4.15% in the composite where the extraction efficiencies of all samples were 4 to 12 times higher than that of commercial silica spin column. The purity of extracted DNA was also evaluated. Purity of DNA samples extracted from commercial silica spin column were between 1.5 - 8.37 value where samples extracted from GO/Cellulose composite were 1.6 - 2.0 value ranges which indicating that extracted DNA is more suitable for further applications. On the other hands, magnetic particles have been developed for purification of genomic or plasmid DNA from different biological sources since it can provide decent amount of DNA extraction in direct application with rapid recovery. Hence, in chapter 5, GO/Cellulose/Magnetic composites were prepared. The mass % of Graphite oxide was keeped at 4.15% and concentration of magnetite inside the composite ranged between 0.2 – 3.98 wt. %. The extraction efficiency was 150 times higher than that of commercial magnetic beads, while the purity of DNA was ranged between 1.7-1.8 value ranges. Finally, chapter 6 summarized the performance and extraction efficiency of GO/Cellulose composite on each type of DNA samples. The results demonstrated that it might be a promising and efficient solid phase material for DNA extraction applications. In this study, GO/Cellulose composite is one of the first researches in the field of applying carbon based material for DNA extraction. Compare to the most common DNA extraction material, silica films, GO/cellulose composite showed higher DNA binding capacity and extraction yield with a simple and environment friendly production. The results observed that (1) It was found that the binding capacity of DNA increased with increasing weight percentage of GO inside the composite, (2) GO/Cellulose composite showed high extraction efficiency not only for ssDNA, but also for Genomic DNA samples, and (3) GO/Cellulose/Magnetite composite exhibits superior extraction efficiency compared to that of commercial magnetic bead product.
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