Detection of K-ras exon 1 mutations by constant denaturant capillary electrophoresis

Biomedical Chromatography : BMC
Chunxia ZhaoQing Yang

Abstract

Among various mutation detection methods, constant denaturant capillary electrophoresis (CDCE) is one of the most common techniques for rapid identification 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 difficulty. 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.

Citations

Sep 20, 2008·Virchows Archiv : an International Journal of Pathology·J H J M van KriekenA Ensari
Jul 17, 2007·Pathology, Research and Practice·Angela PoehlmannRegine Schneider-Stock
Jun 4, 2005·Electrophoresis·Jens Bjørheim, Per Olaf Ekstrøm
Nov 2, 2007·Chemical Reviews·Karel Kleparník, Petr Bocek

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