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What is the difference in sensitivity when using an intercalating dye (e.g., SYBR® Green) vs. a 5’ nuclease assay probe (e.g., PrimeTime® qPCR Assays) to detect PCR product?
Probe-based assays and SYBR assays both have the potential to detect a single copy of target, and thus, can be equally sensitive. However, it is important to remember that SYBR assay fluorescent signal can be generated as a sum of fluorescence due to dimers and other nonspecific products in addition to the desired target. Hence, when using SYBR assays, it is necessary to determine whether or not the fluorescent signal comes entirely from detection of the desired product. Find out more about probe- and intercalating dye assays by visiting our PrimeTime qPCR Assays webpage.