Detection of Alu insertion polymorphisms by novel fluorescent multiplex PCR

• Main Authors: Cheng-Hsien, Wu, 吳政憲
• Other Authors: James chun-I Lee
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/8ry3cm

碩士 === 中央警察大學 === 鑑識科學研究所 === 90 === Alu insertions are short interspersed repetitive elements (SINEs), and have been accumulating in the human genome throughout primate evolution, reaching a copy number of over a million per genome. Each full-length Alu insertion is a dimeric, ~300bp retroposon that is homologous to the 7SL RNA component of the signal-recognition particle. Alu insertion polymorphisms are ideal markers for human evolutionary and individual identification studies because retroposition produces infrequent, irreversible, widely distributed insertion events, each with a known ancestral state. This thesis reports an analysis of 10 polymorphic loci (ACE, B65, HS C3N1, HS 3.23, HS 4.32, PLAT, Ya5 NBC35, Ya5 NBC61, Ya5 NBC148, Ya5 NBC242) in 99 individuals form Taiwanese Han population. Here also develop a novel fluorescence multiplex PCR system in Alu insertion detection and compare it’s power of discrimination to tranditional PCR analysis system. The specific primers were designed to amplify Alu insertion at the side sequence of Alu sequence and intra-Alu insertion sequence. The multiplex PCR product was analyzed by using the polymer capillary electrophoresis of ABI PRISM 310 Genetic Analyzer at 15 kV for 24 minutes at 60˚C. This novel fluorescent multiplex PCR system have reached combined discrimination power to 0.9546. This study indicated that the novel multiplex PCR could be an efficient tool to detection Alu insertion for human genetic diversity, individual identification, paternity determination and forensic analysis. Keywords: Alu insertion, SINEs, polymorphism, multiplex PCR, individual identification, discrimination power.