| Summary: | 碩士 === 國立臺灣大學 === 化學工程學研究所 === 91 === To set up a microarray system tailored to the detection of a specific gene to elucidate the physiological phenomena of the plants, a glass slide was selected as a support for immobilization of DNA. The preliminary experiment was to set up a DNA array for detection of 20mer oligonucleotides with Cy-5 fluorescent labeled to the 5’ end. By measuring contact angle and surface modification of the slide, covalent binding with aldehyde group was superior to the poly-L-lysine immobilized in term of reproducibility and binding force of probe to the glass slide. The former binding method was more toleralent to long period and high salinity conditions. For hybridization of short nucleotide sequence (20mers), one base-mismatch experiment can be completed in two hour reaction.
The microarray of rol genes existed in T-DNA plasmid of transformed root was investigated. Through the experimental works of extracting RNA, fluorescent target labeling by reverse transcription and hybridization with 50mers oligonucleotide-based microarray, it was found the existence and expression of the fragments of rol gene.
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