Multiplex PCR and capillary gel electrophoresis for analysis of gene deletion and duplication types of Duchenne muscular dystrophy

• Main Authors: Chung-An Chen, 陳俊安
• Other Authors: Shou-Mei Wu
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/04162802398284553996

碩士 === 高雄醫學大學 === 藥學研究所 === 99 === Duchenne muscular dystrophy (DMD) is an X-linked genetic disease which is the most serious type in the muscular dystrophy diseases. Due to the lack of protein production, the muscle cells of DMD patients become weak and fragile. Till now, there is none effective treatment. DMD is affected by the largest gene in human, the related gene mutations are complicated. In this study, one genotyping method was established for analysis of DMD gene deletions and duplications in exon 44 to 55. CYBB and pyruvate kinase exon 10 (PK exon 10) are used as internal standards. PCR conditions were optimized for multiplex amplifications. Capillary gel electrophoresis (CGE) was used for simultaneous determination of 12 exons in DMD. The separation conditions was performed by 1.2% PEO in 1X TBE buffer at －6 kV and 25℃. After validation, this PCR-CGE method was applied for DMD patients. Furthermore, this method will be used for eugenics in clinical applications.