PCR ReadyMix Works for PCR but Fails in qPCR (PCR (Sigma Guide))

Question

Accepted Answer

ReadyMix produces amplification in standard PCR but completely fails in real-time qPCR applications; equivalent products from other suppliers work well Common causes: 1) REDTaq ReadyMix was used for real-time qPCR (red dye interferes with fluorescence detection) 2) ReadyMix without reference dye was used on instrument requiring reference dye or incorrect concentration was included 3) ReadyMix designed for use with probes was used without including a probe in the reaction Solutions: 1) Use appropriate ReadyMixes for the desired PCR application 2) Ensure red dye products are not used for qPCR as they interfere with fluorescence detection in most qPCR instruments 3) Verify ReadyMix choice matches instrument requirements and application type