1. What Is qPCR Efficiency?

qPCR efficiency refers to the amplification efficiency of the polymerase chain reaction. It indicates how well the target DNA is being amplified during each cycle of the PCR process. An efficiency of 100% means the amount of product doubles exactly with each cycle.

2. How Does the Calculator Work?

The calculator uses the qPCR efficiency equation:

Where:

• \( slope \) — The slope of the standard curve (dimensionless)

Explanation: The equation converts the slope of the standard curve (typically from a plot of Cq values vs. log input DNA) into a percentage efficiency value.

3. Importance of qPCR Efficiency

Details: Knowing the PCR efficiency is essential for accurate quantification of gene expression. Efficiency values between 90-110% are generally considered acceptable, with 100% being ideal.

4. Using the Calculator

Tips: Enter the slope value from your standard curve. The slope can be obtained from your qPCR instrument's software when creating a standard curve plot.

5. Frequently Asked Questions (FAQ)

Q1: What is an ideal qPCR efficiency?
A: The ideal efficiency is 100%, meaning the DNA doubles perfectly each cycle. Acceptable range is typically 90-110%.

Q2: What does it mean if my efficiency is below 90%?
A: Low efficiency suggests inhibition or suboptimal reaction conditions. You may need to optimize primer concentrations or reaction conditions.

Q3: What if my efficiency is above 110%?
A: High efficiency may indicate primer-dimer formation or contamination. Check your melt curves and no-template controls.

Q4: How do I get the slope value?
A: The slope is generated when you create a standard curve plot in your qPCR software, plotting Cq values against the log of the input DNA quantity.

Q5: Can I use this for absolute quantification?
A: Yes, knowing your reaction efficiency is crucial for accurate absolute quantification in qPCR experiments.