Capillary electrophoresis of double-stranded DNA fragments using a new fluorescence intercalating dye EvaGreen.
EvaGreen is a new DNA intercalating dye successfully used in quantitative real-time PCR. In the present work, we firstly apply EvaGreen to the analysis of dsDNA by CE with LIF detection. Comparisons of EvaGreen dye with the commonly used dyes SYBR Green I and SYBR Gold were preformed in dsDNA analysis by CE. The linear range of dsDNA using EvaGreen was slightly wider than that using SYBR Gold and SYBR Green I, and the detection limits of dsDNA were not significantly different for the three dyes. Good separations of dsDNA fragments were obtained using the three dyes. Reproducibility of migration time and the peak area of dsDNA fragments with EvaGreen were better than those for SYBR Green I and SYBR Gold. The RSD values were 0.24-0.27% for migration time and 3.45-7.59% for peak area within the same day, 1.35-1.63% for migration time and 6.72-12.05% for peak area for three days. Our data demonstrated that EvaGreen is well suited for the dsDNA analysis by CE with LIF detection.
Sang
Fuming
F
College of Chemistry and Chemical Engineering, Shanghai Jiaotong University, Shanghai, P. R. China.
Ren
Jicun
J
D052160:Benzothiazoles; D004247:DNA / Q000032:analysis*; D053938:DNA Fragmentation; D003959:Diamines; D019075:Electrophoresis, Capillary / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry*; D008422:Materials Testing; D009930:Organic Chemicals / Q000737:chemistry; D011804:Quinolines; D012680:Sensitivity and Specificity
D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA