qPCR Efficiency Calculator
What is qPCR Efficiency?
qPCR efficiency is a crucial metric that indicates the performance of a PCR reaction. It represents the rate at which a PCR amplicon is generated, specifically the multiplication factor per cycle. In an ideal PCR reaction, the amount of template doubles during each cycle, corresponding to an efficiency of 2.0 or 100%. However, in real-world conditions, various factors can affect PCR efficiency, including primer design, template quality, and reaction conditions. Understanding and monitoring PCR efficiency is essential for accurate quantification and reliable experimental results.
How to Use qPCR Efficiency Calculator
Step 1: Obtain your standard curve slope value from your qPCR experiment. This is typically generated by plotting Ct values against log template concentration.
Step 2: Enter the slope value in the calculator. The slope should be negative and typically falls between -3.1 and -3.6 for optimal reactions.
Step 3: Optionally, enter your R² value if you want to assess the quality of your standard curve. R² values should be >0.98 for reliable results.
Step 4: Click the calculate button to obtain your results. The calculator will display both the efficiency value and the percentage efficiency.
Step 5: Interpret your results using the provided explanation. For optimal qPCR reactions, aim for efficiency between 90-110% (1.90-2.10).
Step 6: If your efficiency falls outside the optimal range, consider reviewing your experimental setup, including primer design, template quality, and reaction conditions.