1. What is qPCR Efficiency?

qPCR efficiency describes how effectively the PCR reaction amplifies the target sequence. An ideal efficiency is 100%, meaning the amount of product doubles with each cycle.

2. How Does the Calculator Work?

The calculator uses the qPCR efficiency equation:

Where:

• \( slope \) — Slope of the standard curve (dimensionless)
• \( eff \) — PCR efficiency (%)

Explanation: The equation converts the standard curve slope into a percentage efficiency value.

3. Importance of Efficiency Calculation

Details: Knowing your qPCR efficiency is crucial for accurate quantification, as it affects the calculation of fold changes and absolute quantities.

4. Using the Calculator

Tips: Enter the slope value from your standard curve plot. The slope should be negative (typically between -3.1 to -3.6 for good efficiency).

5. Frequently Asked Questions (FAQ)

Q1: What is an ideal qPCR efficiency?
A: 90-110% is generally acceptable, with 100% being perfect. Below 90% or above 110% may indicate problems.

Q2: How do I get the slope value?
A: The slope comes from plotting Ct values against the log10 of template concentration in your standard curve.

Q3: Why is efficiency important?
A: Efficiency affects quantification accuracy. Reactions with different efficiencies cannot be directly compared.

Q4: What causes poor efficiency?
A: Common causes include poor primer design, inhibitor presence, pipetting errors, or suboptimal reaction conditions.

Q5: Can I use this for absolute quantification?
A: Yes, but only if your standard curve and samples have similar efficiencies.