1. What is a qPCR Amplification Efficiency Calculator?

Definition: This calculator determines the amplification efficiency of a qPCR reaction based on the slope of the standard curve.

Purpose: It helps molecular biologists assess the performance of their qPCR assays and ensure accurate quantification.

2. How Does the Calculator Work?

The calculator uses the formula:

Where:

• \( Eff \) — Amplification efficiency (decimal)
• \( slope \) — Slope of the standard curve (dimensionless)

Explanation: The efficiency is derived from the slope of the standard curve plot (Ct vs. log template quantity).

3. Importance of qPCR Efficiency Calculation

Details: Proper efficiency calculation ensures accurate quantification, with ideal efficiency being 1 (or 100%).

4. Using the Calculator

Tips: Enter the slope value from your standard curve. The calculator provides both decimal and percentage efficiency values.

5. Frequently Asked Questions (FAQ)

Q1: What is an ideal efficiency value?
A: The ideal efficiency is 1 (or 100%), meaning the DNA doubles each cycle. Acceptable range is typically 0.9-1.1 (90-110%).

Q2: How do I get the slope value?
A: The slope is obtained from the standard curve plot generated by your qPCR software (Ct vs. log template concentration).

Q3: What does efficiency > 1 mean?
A: Efficiency > 1 (100%) suggests possible PCR artifacts like primer-dimer formation or inaccurate standard curve.

Q4: What does efficiency < 1 mean?
A: Efficiency < 1 suggests inhibition or suboptimal reaction conditions.

Q5: How does efficiency affect quantification?
A: The efficiency value is used in quantification algorithms to calculate the starting quantity of template.