1. What is QPCR Efficiency?

QPCR efficiency refers to the percentage of template DNA that is amplified in each cycle of the PCR reaction. It is a critical parameter for accurate quantification in real-time PCR experiments.

2. How Does the Calculator Work?

The calculator uses the QPCR efficiency equation:

Where:

• \( E \) — PCR efficiency (expressed as percentage)
• \( S \) — Slope of the standard curve (Ct vs log concentration)

Explanation: The equation calculates the amplification efficiency based on the slope of the standard curve. An ideal efficiency is 100%, meaning the DNA doubles every cycle.

3. Importance of Efficiency Calculation

Details: Accurate efficiency calculation is crucial for reliable quantification in qPCR experiments. It affects the accuracy of relative quantification and absolute copy number determination.

4. Using the Calculator

Tips: Enter the slope value from your standard curve. The slope should be obtained from a plot of Ct values versus the logarithm of template concentration.

5. Frequently Asked Questions (FAQ)

Q1: What is the ideal qPCR efficiency?
A: The ideal efficiency is 100%, which corresponds to a slope of -3.32. Efficiency between 90-110% is generally considered acceptable.

Q2: Why is efficiency important in qPCR?
A: Efficiency affects the accuracy of quantification. Variations in efficiency can lead to errors in calculating fold changes or absolute copy numbers.

Q3: How do I obtain the slope value?
A: The slope is obtained from the standard curve generated by plotting Ct values against the logarithm of known template concentrations.

Q4: What factors affect qPCR efficiency?
A: Primer design, template quality, reaction conditions, inhibitor presence, and instrument calibration can all affect efficiency.

Q5: Can I use this calculator for any qPCR experiment?
A: Yes, this calculator can be used for any qPCR experiment where you have generated a standard curve and obtained the slope value.