We recommend using SYBR® Green PCR Master Mix from Applied Biosystems.
1. Mix the gene-specific forward and reverse primer pair. Each primer (forward or reverse) concentration in the mixture is 5 pmol/pL.
2. A real-time PCR reaction volume can be either 50 pL or 25 pL. Prepare the following mixture in each optical tube. Do not use more than 1 pL cDNA template.
3. Set up the experiment on a real-time PCR machine with the following PCR program.
3) 95°C 15 seconds ^ 60°C 30 seconds ^ 72°C 30 seconds, 40 cycles
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