Effect of polymer matrix and glycerol on rapid single-strand conformation polymorphism analysis by capillary and microchip electrophoresis for detection of mutations in K-ras gene

被引:23
作者
Endo, Y
Zhang, L
Katashima, R
Itakura, M
Doherty, EAS
Barron, AE
Baba, Y
机构
[1] Nagoya Univ, Grad Sch Engn, Dept Appl Chem, Nagoya, Aichi 4648603, Japan
[2] Univ Tokushima, Grad Sch Pharmaceut Sci, Dept Mol & Pharmaceut Biotechnol, Tokushima 770, Japan
[3] CREST, Japan Sci & Technol Corp, Chiba, Japan
[4] Furuno Elect Co Ltd, Nishinomiya, Hyogo, Japan
[5] Univ Tokushima, Inst Gen Res, Tokushima 770, Japan
[6] Northwestern Univ, Dept Chem & Biol Engn, Evanston, IL USA
[7] Natl Inst Adv Ind Sci & Technol, Hlth Technol Res Ctr, Takamatsu, Kagawa, Japan
[8] Japan Biol Informat Consortium, Genet Diversificat Anal Project, Sect Diabet, Genotyping Div, Tokushima, Japan
[9] Appl Biosyst Inc, Tokyo, Japan
关键词
capillary electrophoresis; microchip electrophoresis; mutation detection; single-strand conformation polymorphism;
D O I
10.1002/elps.200500142
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
We present the rapid single-strand conformation polymorphism (SSCP) analysis by capillary and microchip electrophoresis to detect the mutations in K-ras gene. Parameters that might affect the analysis of mutation in K-ras gene, such as the polymer and the additive in the sieving matrix, have been studied systematically. Under the optimal conditions, the analysis of seven mutants of K-ras gene could be finished within 10 min by capillary electrophoresis (CE). Furthermore, with the wild-type gene as the inner standard, the analysis accuracy of mutations could be improved. In addition, by studying the properties of polymer solutions, the matrix suitable for microchip electrophoresis was found, and the detection of mutations in K-ras gene could be further shortened to 1 min.
引用
收藏
页码:3380 / 3386
页数:7
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