Detection of DNA concentration in PCR using QDs

• Main Authors: Jheng-Yi Lin, 林政逸
• Other Authors: Yaw-Jen Chang
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/77463912705378864835

碩士 === 中原大學 === 機械工程研究所 === 94 === Abstract Due to the rapid development of nano technology in recent years, nano-particles have been widely used in a variety of research areas, especially in the biomedical science. The miniaturization in matter scale brings huge changes in matter’s physical and chemical properties because of the quantum effect. It may also lead to new characteristics such as optical and electrical properties. This thesis is concerned with the measurement of DNA concentration in the polymerase chain reaction (PCR) by means of the specific optical property of nano-particles. Particularly, the quantum dots were adopted as the fluorescent material for the detection of DNA concentration. The merits of quantum dots’ properties include narrow fluorescent wavelength, wild excited wavelength, and long fluorescent time. In addition, the color of light emitted from the quantum dot is determined by its size. After coating, it can be used in vivo. For the biological treatment, the surface property of quantum dots was firstly changed from hydrophobic to hydrophilic by coating. Then, the quantum dots that have specific functional groups can synthesize with the modified primers. By this procedure, the primers carry the fluorescent material, i.e. the quantum dots. After the pretreatment, the polymerase chain reactions were performed. Furthermore, the fluorescent change was observed by the electrophoresis process so that the relationship between the DNA concentration and the fluorescent intensity of quantum dots can be determined. The results of experiment show that DNA cloning is successful but not complete. Quantum dots would be affected by high temperature and caused the fluorescent intensity decreasing significantly. We will adjust the recipe of quantum dots surface modification and bioconjugation method to improve the fluorescent intensity and DNA cloning efficiency in the future.