高分辨率熔解曲线(HRM)是基于新型饱和荧光染料的基因分型技术,本实验旨在研究荧光染料类型、添加量以及添加方式对HRM分型效果的影响。实验分析了3种荧光染料(LC Green、JJ Green、SYBR Green I)、不同荧光染料浓度、染料添加方式对HR...高分辨率熔解曲线(HRM)是基于新型饱和荧光染料的基因分型技术,本实验旨在研究荧光染料类型、添加量以及添加方式对HRM分型效果的影响。实验分析了3种荧光染料(LC Green、JJ Green、SYBR Green I)、不同荧光染料浓度、染料添加方式对HRM分型差异的比较。结果表明:3个染料中LC Green效果最佳,JJ Green次之,而SYBR Green I不及前两种;染料添加量会对HRM分型产生较大的影响,染料添加量为0.8μL时为最优反应量;而染料添加方式对分析结果没有影响。展开更多
Terbium （Tb） has been extensively used as a fluorescence probe for the identification of calcium-binding sites in proteins and for fluorometric analysis of organic ligands. In the current study, we reported that HEP...Terbium （Tb） has been extensively used as a fluorescence probe for the identification of calcium-binding sites in proteins and for fluorometric analysis of organic ligands. In the current study, we reported that HEPES, a commonly used pH buffer reagent, significantly enhanced the characteristic emission of Tb at 585 nm. The maximum emission of Tb at 490 nm and 549 um were also enhanced by HEPES to a less extent. Thus, cautions should be taken when quantitative analysis is performed based on the fluorescence emission of Tb at 549 nm, since the emission may vary due to the buffer reagents. Additionally, the fluorescence intensity at 585 um was proportional to the concentration of both HEPES and terbium ions, which might be utilized to develop new fluorometric analytical methods.展开更多
文摘高分辨率熔解曲线(HRM)是基于新型饱和荧光染料的基因分型技术,本实验旨在研究荧光染料类型、添加量以及添加方式对HRM分型效果的影响。实验分析了3种荧光染料(LC Green、JJ Green、SYBR Green I)、不同荧光染料浓度、染料添加方式对HRM分型差异的比较。结果表明:3个染料中LC Green效果最佳,JJ Green次之,而SYBR Green I不及前两种;染料添加量会对HRM分型产生较大的影响,染料添加量为0.8μL时为最优反应量;而染料添加方式对分析结果没有影响。
基金National Natural Science Foundation of China(Grant No.20637010 and 20871008).
文摘Terbium （Tb） has been extensively used as a fluorescence probe for the identification of calcium-binding sites in proteins and for fluorometric analysis of organic ligands. In the current study, we reported that HEPES, a commonly used pH buffer reagent, significantly enhanced the characteristic emission of Tb at 585 nm. The maximum emission of Tb at 490 nm and 549 um were also enhanced by HEPES to a less extent. Thus, cautions should be taken when quantitative analysis is performed based on the fluorescence emission of Tb at 549 nm, since the emission may vary due to the buffer reagents. Additionally, the fluorescence intensity at 585 um was proportional to the concentration of both HEPES and terbium ions, which might be utilized to develop new fluorometric analytical methods.
