Premium
Detection of K‐ras exon 1 mutations by constant denaturant capillary electrophoresis
Author(s) -
Zhao Chunxia,
Xu Guowang,
Shi Xianzhe,
Ma Jianmei,
Lu Shen,
Yang Qing
Publication year - 2004
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.353
Subject(s) - chemistry , capillary electrophoresis , chromatography , electrophoresis , capillary action , constant (computer programming) , exon , biochemistry , gene , thermodynamics , physics , computer science , programming language
Among various mutation detection methods, constant denaturant capillary electrophoresis (CDCE) is one of the most common techniques for rapid identication of known or unknown mutations. In this report, a CDCE analysis method with homemade linear polyacrylamide (LPA) kit was developed on ABI 310 genetic analyzer, the effect and relationship of various denaturing factors in CDCE analysis were investigated and K‐ras gene mutations of 31 coloerctal cancer patients were detected. Results indicate that, with the increase of chemical danaturant concentration, the optimum temperature was lowered, and when the concentration of urea (formamide) was higher than 7 m (40%), the homoduplex and heteroduplex of mutant samples were separated with difculty. Detection results of K‐ras gene in colorectal samples indicated that mutations were present in eight (26%) of 31 patients; most mutations were localized in codon 12, which is thought to be a critical step and plays an important role in human colorectal carcinogenesisas. Copyright © 2004 John Wiley & Sons, Ltd.