Experimental Conditions of Detecting rs2268388 Polymorphism of ACACB Gene

［Abstract］Objective To establish a rapid and specific quantitative PCR assay for detecting of ACACB gene rs2268388 polymorphism. Methods The primers and Taqman probe targeted at signature sequence of ACACB gene rs2268388 polymorphism were designed and applied to real time PCR assays. The different concentrations were set up， and the sensitivity and specificity were evaluated. Results Initial denaturation time of PCR was 10 min， annealing and extending time were 1 min， probe concentration of Taqman PCR was 0.5 μL（0.2 μM），TaqMan Genotyping Master Mix was 5 μL， dH2O was 2.5 μL，and total volume was 10 μL. At this condition，the result of ACACB gene rs2268388 polymorphism showed the high specificity of the reaction. Conclusion The assay was sensitive and specific for rapid identification and quantitative detection of ACACB gene rs2268388 polymorphism. It will be an effective method for the surveillance and diagnosis of ACACB gene rs2268388 polymorphism.