Capillary Electrophoresis Using Single Strand Conformation Polymorphism for the Analysis of Human Obesity DNA

Journal of the Korean Chemical Society

Sangmin Jang¹  Seong Ho Kang¹  Ho-Keun Yi¹ 

关键词: Human Obesity DNA;    Polymerase Chain Reaction;    Single Strand Conformation Polymorphism/Capillary Electrophoresis;    Point Mutation;   

DOI  :  

学科分类：化学（综合）

来源: Korean Chemical Society

【 摘 要 】

We have investigated a capillary electrophoresis (CE) method using single strand conformation polymorphism (SSCP) for the rapid detection of human obesity DNA. Using polymerase chain reaction (PCR) we amplified human obesity DNA, Leu72Met mutation made an estimate of child obesity DNA mutant. The ss-DNA fragments, which are different to original DNA fragments, were prepared by directly immersing in 1 oC ice bath after denaturing the amplified 207-bp human obesity ds-DNA sample at 95 oC. The prepared ss-DNA fragments were separated in a fused-silica capillary (50 mm I.D. ¡¿365 mm O.D. ¡¿60 cm total length; 30 cm effective length) filled with 1.5% poly (vinylpyrrolidone) (Mr 1 300 000), 1.0% poly(ethyleneoxide) (Mr 600 000) and 5%(w/w) glycerol as the sieving matrix and running buffer, based on gel electrophoretic separation. Under the electric field of 375 V/cm, the human obesity DNA was analyzed within 9 min at the CE. The SSCP-CE showed at least the 10-times faster analysis time than the traditional slab gel electrophoresis.

【 授权许可】

Unknown