摘要
以荧光染料Fam和Joe分别标记野生型和突变型双链探针作为均相检测探针,以构建的DNA模板作为研究模型,采用固定波长差同步荧光分析法对PCR反应产物进行终点检测。通过对HFE基因C282Y点突变的检测,并以限制性内切核酸酶RsaI证实,该方法是一种廉价、快速、可靠的筛查遗传性血色病基因C282Y突变的方法,该法可扩展到各种基因的突变检测。
We described in the paper a new highthroughput screening method for Cys282Tyr mutation in hereditary haemochromatosis with doublestranded probe using synchronous fluorometry.The probe for wild type was labeled with Fam,the probe for mutant type was labeled with Joe.After PCR,reaction tubes were transferred to a spectrofluorometer,where synchronous spectra were scanned in a constantwavelength mode.The genotype could be obtained through the appearance of the fluorescence peaks corresponding to each probe.The results were totally in agreement with restriction endonuclease analysis.Considering the simplicity,low cost and specificity,this approach could be generally applied to detect varieties of gene mutations.
出处
《遗传》
CAS
CSCD
北大核心
2003年第1期9-13,共5页
Hereditas(Beijing)
基金
国家自然科学基金项目(30170834)资助