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Validation of RT-qPCR to determine primer efficiency. To validate our primers, we conducted a standard curve experiment using 5x step dilution starting with 20 ng. Standard curves for the RT-qPCR primer validation were created by 5x step-wise dilution of total RNA from midbrain samples containing 20 ng to about 6 pg. The experiment was performed in triplicate. Data is presented as mean ± SEM. All primers passed the validation with acceptable efficiencies.
