Genotyping of K-ras codons 12 and 13 mutations in colorectal cancer by capillary electrophoresis.

摘要: Abstract Point mutations of the K - ras gene located in codons 12 and 13 cause poor responses to anti-epidermal growth factor receptor (anti-EGFR) therapy colorectal cancer (CRC) patients. Besides, have also been proven play an important role human tumor progression. We established a simple effective capillary electrophoresis (CE) method for simultaneous point mutation detection gene. combined one universal fluorescence-based nonhuman-sequence primer two fragment-oriented primers tube, performed this two-in-one polymerase chain reaction (PCR). PCR fragments included wild type seven at The amplicons were analyzed by single-strand conformation polymorphism (SSCP)-CE method. CE analysis was using 1× Tris–borate–EDTA (TBE) buffer containing 1.5% (w/v) hydroxyethylcellulose (HEC) (MW 250 000) under reverse polarity with 15 °C 30 °C. Ninety patients blindly genotyped developed results showed good agreement those DNA sequencing SSCP-CE feasible screening populations.