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Design of PCR Primers: Helpful Web Sites that Contain Tips, Software, and Sequences42. 3.1.3. Using the Primers in PCRs. 1.1 Key Concepts of Real-Time PCR. 2. 1.1.1 What Is Real-Time PCR? 2. 1.1.2 How Real-Time PCR Works. 3. 1.1.3 Hallmarks of an Optimized qPCR Assay. SuperScript™ III Platinum® One-Step Quantitative RT-PCR System. Ref: Invitrogen 1732-020. Primers and probes. Name. Sequences (5'-3'). Length. (bases). PCR.The polymerase chain reaction (PCR) is one of the most powerful technologies in molecular biology. Using PCR, specific sequences within a DNA or