荧光染料SYBR GreenI及EB在定量PCR中敏感性比较

Sensitivity comparison of SYBR Green I with EB in Q-PCR

目的:比较荧光染料SYBRGreenI及EB在定量PCR诊断中检测DNA的敏感性。方法:分别用琼脂糖凝胶电泳SYBRGreenI染色及EB染色,检测26例21三体综合征患者及20例正常人DNAPCR扩增产物,并进行光密度分析比较。结果:SYBRGreenI染色于24个循环PCR产物带型清楚,可准确定量检测;EB染色则在28个循环才能进行定量分析。结论:荧光染料SYBRGreenI染色较EB染色测量DNA敏感、快捷,在定量PCR技术中值得推广应用。

Aim: To compare the sensitivity of SYBR Green I stain with ethidium bromide(EB)stain in quantitative PCR(Q-PCR).Methods: The DNA samples were isolated from 26 cases of Down's syndrome and 20 normal subjects. The DNA amplification products were subjected to agarose gels with SYBR Green I stain and EB stain, respectively. The optical densitometry of the PCR products was measured.Results: DNA amplification products in 24 cycles stained with SYBR Green I were clearly visualized and quantitatively detected,but DNA amplification products stained with EB could not be quantitatively analyzed until 28 cycles.Conclusion: SYBR Green I is more sensitive and rapid for detecting DNA in Q-PCR assay compared with EB.