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Does the PrimeTime Gene Expression Master Mix use a hot-start polymerase?
Yes, PrimeTime™ Gene Expression Master Mix uses an antibody-mediated, hot-start polymerase. During the initial extended denaturation step of qPCR cycling, polymerase activity is unblocked when the antibody releases from the polymerase. This "hot-start" DNA polymerase enhances specificity and sensitivity of the qPCR by avoiding nonspecific amplification of DNA and primer dimer formation. Therefore, it is critical to follow the cycling conditions provided in the PrimeTime Gene Expression Master Mix protocol.