JumpStart Taq ReadyMix Underperforming Due to Incorrect Hot Start Protocol (PCR (Sigma Guide))

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JumpStart Taq ReadyMix does not work as well as similar products from different suppliers; amplification is weak or absent despite correct assay design Common causes: 1) Incorrect initial incubation of 95°C for 15 min was used (standard Hot Start protocol for chemically inactivated enzymes inappropriately applied to antibody-inactivated enzyme) 2) Primer optimization conditions differ slightly between ReadyMixes 3) Extended incubation time partially inactivates JumpStart Taq enzyme Solutions: 1) Use initial incubation of 3 min at 94°C to activate JumpStart Taq (specific protocol for antibody-inactivated enzyme) 2) Adjust primer concentration or annealing temperature (Ta) to optimize for the specific ReadyMix 3) Avoid longer incubations that will partially inactivate the enzyme