Comparison of fluorescent intercalating dyes for quantitative loop-mediated isothermal amplification (qLAMP)

Abstract

Real-time or quantitative loop-mediated isothermal amplification (qLAMP) is a promising technique for the accurate detection of pathogens in organisms and the environment. Here we present a comparative study of the performance of six fluorescent intercalating dyes-SYTO-9, SYTO-13, SYTO-82, SYBR Green I, SYBR Gold, EvaGreen-in three different qLAMP model systems. SYTO-9 and SYTO-82, which had the best results, were used for additional enzyme and template titration studies. SYTO-82 demonstrated the best combination of time-to-threshold (Tt) and signal-to-noise ratio (SNR).

Keywords: Bst DNA polymerase; EvaGreen; LAMP; SYBR Gold; SYBR Green I; SYTO-13; SYTO-82; SYTO-9; isothermal amplification.