Preliminary studies on in-situ genome sequencing by chromosome microdissection

• Main Authors: Yi-HuiTsai, 蔡儀慧
• Other Authors: Song-Bin Chang
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/68202670827088891494

碩士 === 國立成功大學 === 生命科學系 === 102 === Genomiphi amplification is one of the whole genome amplification (WGA) tools. It work on generating microgram quantities from low copy number DNA or degraded DNA with multiple displacement amplification (MDA) mechanisms of phi29 DNA polymerase. We described an innovation that using microdissection technique combined illustraTM GenomiPhi Amplification kit (GE Healthcare) to solve the obstacles of conventional sequencing in spending time to construct gene library and genetic map. In order to evaluate the feasibility of this strategy, we chose long arm region of tomato chromosome 6 as material for needle-microdissection and reconfirmed WGA-DNA (dissected segment, 6L) by fluorescence in situ hybridization (FISH). According to the results of 6L and 45S rDNA show the same loci, which means that is not expected results. Therefore, the procedure of this strategy needs to be re-examined. We hope improved technique can get more accuracy and convenient for genome sequencing.