1 year ago
What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.
It depends on several factors. First, try to determine how much reaction you want to run. The primary concentration of your qPCR reaction can range from 15 to 30 microliters. The qPCR machine is also can affect it. For example, MIC real-time PCR apparatus enables you to run 10 microliters of your reaction, consequently, you can save a considerable amount of input chemicals per run. Corbett rotor gene 6000 real-time PCR is also working well with a lower concentration of your qPCR reaction. A final concentration of 200 nM per primer is effective for most reactions. Optimal results may require a titration of primer concentrations between 100 and 500 nM. For dye-based qPCR and Real-time PCR the optimum dose of each R/F primers is mentioned in their protocol and you can select the best concentration for your goal. Before running all samples, try to set up your reaction with some primary runs and using different concentrations of R/F primers. Primer design is also a quintessential step in your project. If you are not an expert in molecular biology, somebody other may support you for this case.
Answered 10 months ago
Share your thoughts or question with experts in your field