1. What is qPCR Efficiency?

qPCR efficiency measures how effectively the DNA template is amplified during each cycle of the PCR reaction. It's crucial for accurate quantification in water quality analysis.

2. How Does the Calculator Work?

The calculator uses the efficiency formula:

Where:

• \( slope \) — Slope value from standard curve

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

3. Importance of Efficiency Calculation

Details: Proper efficiency (90-110%) ensures accurate quantification of target DNA in water samples. Deviations may indicate PCR inhibition or primer issues.

4. Using the Calculator

Tips: Enter the slope value from your standard curve. Ideal slope for water samples is typically between -3.1 to -3.6.

5. Frequently Asked Questions (FAQ)

Q1: What is the ideal efficiency range?
A: 90-110% is generally acceptable for water analysis. Values outside this range may require optimization.

Q2: How does water quality affect qPCR efficiency?
A: Water contaminants like humic acids can inhibit PCR, lowering efficiency.

Q3: When should efficiency be measured?
A: With every qPCR run, using a standard curve with known DNA concentrations.

Q4: What causes low efficiency in water samples?
A: Common causes include PCR inhibitors in water, poor primer design, or suboptimal reaction conditions.

Q5: How to improve efficiency for water samples?
A: Sample filtration, dilution, or adding PCR facilitators like BSA can help.