The Study of Detection of Environmental Microorganisms by Different DNA Labeling Methods

• Main Authors: Chia-Chen Wang, 王家振
• Other Authors: Yu-Jan Chou
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/ve3u42

碩士 === 輔英科技大學 === 生物技術系碩士班 === 96 === In EMSA(electrophoresis mobility shift assay, EMSA)、northern blotting and in-situ hybridization experimental procedure, biotinylated oligo are often labeled in oligonucleotide probes. Due to the enzyme reaction between biotin and streptavidin-gold, labeled DNA were observed. The genomic 16S rDNA of Acidovorax sp. were extracted from the Anoxic-Oxic by biological nitrogen removal procedures (the hydraulic retention time = 10 hours, the sludge age = 30 days, the internal reflux ratio = 2.5) which were operated A method(biotin-labeled dUTPs), B method(biotin-labeled forward primer) and C method(digoxigenin-labeled dUTPs) to be a specific sample for strip chip. In order to find the optimal experimental condition, different concentration of specific sample and volume of streptavidin-gold were examination in this research. Moreover, Biocapt software was adapted to analysis the intensity of signal in genomic 16S rDNA of Rhodobacter sp. detection. According to signal intensity, retention time and cost analysis results, this research offered an alternative optimal procedure in environmental bacterium detection.