How does SYBR Green work in qPCR?

Posted June 22, 2020

Answer

SYBR Green is one of the most commonly used fluorescent dyes in qPCR. It binds to double-stranded DNA molecules by intercalating between the DNA bases. Once intercalated to DNA, SYBR Green becomes less mobile, causing its energy to be released as fluorescence. Therefore, the fluorescence intensity is directly associated with the concentration of double-stranded DNA, which can be measured at the end of each amplification cycle to determine the PCR progress.

Additional resources

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*

Postollec, F., Falentin, H., Pavan, S., Combrisson, J., & Sohier, D. (2011). Recent advances in quantitative PCR (qPCR) applications in food microbiology. Food microbiology, 28(5), 848-861.

Valasek, M. A., & Repa, J. J. (2005). The power of real-time PCR. Advances in physiology education, 29(3), 151-159.