qPCR Efficiency Calculator
Calculate amplification efficiency, R², and slope from standard curve data
🧪 Standard Curve Data
Dilution Points
| Dilution | Ct Value |
|---|
📚 qPCR Efficiency Concepts
Efficiency = (10^(-1/slope) – 1) × 100%
Standard formula for qPCR efficiency calculation
Key Parameters:
- • Efficiency: 90-110% is optimal, 100% means perfect doubling
- • R²: >0.99 indicates excellent linearity
- • Slope: -3.32 indicates 100% efficiency
- • Dynamic Range: 5-6 logs for good assays
📈 Efficiency Results
Amplification Efficiency
100.0%
Slope
-3.322
R² Value
1.000
Y-Intercept
40.0
📊 Quality Assessment
Efficiency Status
Optimal
Linearity
Excellent
Dynamic Range
6 logs
Recommendation
Acceptable
🧬 What is qPCR Efficiency?
qPCR efficiency measures how effectively DNA is amplified during each PCR cycle. Perfect efficiency (100%) means the DNA quantity doubles every cycle.
Efficiency is calculated from the standard curve slope using the formula: Efficiency = (10^(-1/slope) – 1) × 100%
Slope = -3.322 → Efficiency = 100%
Ideal slope value for perfect efficiency
⚖️ Quality Parameters
Several parameters determine qPCR assay quality:
- ✓ Efficiency: 90-110% (optimal range)
- ✓ R²: >0.99 (excellent linearity)
- ✓ Slope: -3.1 to -3.6 (acceptable range)
- ✓ Dynamic Range: 5-6 logs (good sensitivity)
- ✓ Y-intercept: Indicates detection limit
🔍 Troubleshooting Tips
Common issues and solutions:
- • Low Efficiency: Check primer design, optimize annealing temperature
- • Poor R²: Verify pipetting accuracy, check template quality
- • High Slope: May indicate inhibition or poor primer efficiency
- • Narrow Range: Extend dilution series, check instrument sensitivity
Note: Always include proper controls and validate with biological replicates.