1. What is PCR Amplification?

PCR (Polymerase Chain Reaction) is a technique used to amplify specific DNA sequences exponentially. Each cycle theoretically doubles the number of DNA copies, allowing detection of minute quantities of DNA.

2. How Does the Calculator Work?

The calculator uses the PCR amplification equation:

Where:

• Initial Copies — Starting number of DNA template molecules
• Cycles — Number of PCR amplification cycles performed
• Final Copies — Estimated total number of DNA copies after amplification

Explanation: The equation models ideal PCR amplification where each cycle perfectly doubles the DNA quantity.

3. Importance of Copy Number Calculation

Details: Understanding PCR amplification helps in experimental design, determining optimal cycle numbers, and ensuring sufficient product for downstream applications.

4. Using the Calculator

Tips: Enter the starting copy number (typically 1-1000 for single-copy genes) and the number of PCR cycles (typically 20-40). The calculator will estimate the final copy number.

5. Frequently Asked Questions (FAQ)

Q1: Is the calculation always accurate?
A: This assumes 100% efficiency (perfect doubling each cycle). Actual efficiency is typically 90-100%, so results may be slightly lower.

Q2: What's a typical initial copy number?
A: For single-copy genes in genomic DNA, it's approximately equal to the number of genome equivalents in your sample.

Q3: Why does the number get so large?
A: PCR is exponential - 30 cycles with 1 starting copy theoretically yields over 1 billion copies (1 × 2³⁰ = 1,073,741,824).

Q4: When does amplification plateau?
A: Typically after 30-40 cycles due to reagent depletion, product reannealing, and enzyme inhibition.

Q5: Can I use this for qPCR?
A: This calculates theoretical maximum, while qPCR accounts for actual amplification efficiency and provides quantification relative to standards.