Comparison of SYTO9 and SYBR Green I for real-time polymerase chain reaction and investigation of the effect of dye concentration on amplification and DNA melting curve analysis.

摘要: Following the initial report of use SYBR Green I for real-time polymerase chain reaction (PCR) in 1997, little attention has been given to development alternative intercalating dyes this application. This is surprising considering reported limitations I, which include limited dye stability, dye-dependent PCR inhibition, and selective detection amplicons during DNA melting curve analysis multiplex PCRs. We have tested an first detailed evaluation SYTO9. Our findings demonstrate that SYTO9 produces highly reproducible curves over a broader range concentrations than does far less inhibitory not appear selectively detect particular amplicons. The low inhibition high reproducibility means it can be readily incorporated into conventional at broad concentrations, allowing closed tube by analysis. These features simplify improved will make useful diagnostic context.