Abstract
Objective To establish a clinically feasible method for detection of BRAF gene V600E mutation.Methods Based on the known sequences of BRAF wild-type and mutation samples,specific polymerase chain reaction and sequencing primers targeting at the BRAF gene 15 exon V600E mutation were designed.This allowed the establishment of SYRB Green PCR-Sanger sequencing technique for the detection of BRAF V600E mutation,for methodological evaluation and assay of 21 colorectal cancer samples.Results The SYRB Green PCR-Sanger sequencing method for detection of BRAF gene V600E mutation was established successfully,which harbored high assay sensitivity (5.0 × 101 copies/μl) and repeatability and acceptable linear range (5× 101 to 5× 107 copies/μl).The mutation rate was 9.5％ (2/21) in 21 colorectal cancer samples.Condnsion The SYRB Green PCR-Sanger sequencing method for detection of BRAF gene V600E mutation is successfully established and can be applied in assessments for clinical samples. Key words: Mutation; Polymerase chain reaction; Oligonucleotide array sequence analysis; Genes, BRAF
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