Normalize RNA samples to 1 ng/μl. 2. Thaw qPCR kit components on ice, mix gently, and centrifuge briefly. 3. Calculate the number of samples, including controls, and set up duplicate PCR reactions for each template/assay combination. 4. Distribute 10 μl of the reaction mix into each 0.2 ml qPCR well. 5. Seal the wells, mix, centrifuge briefly, and run the following program: 50°C (2 min), 95°C (10 min), 95°C (15 sec), 60°C (1 min), for 40 cycles. 6. Analyze data and adjust conditions if results are inconsistent before repeating the qPCR setup.