1. What is PCR Product Calculation?

The PCR product calculation estimates the final amount of DNA after polymerase chain reaction (PCR) amplification based on the initial amount of DNA and the number of amplification cycles.

2. How Does the Calculator Work?

The calculator uses the PCR product formula:

Where:

• Initial Amount — Starting quantity of DNA (unitless)
• Number of Cycles — PCR amplification cycles (unitless)
• Product — Final amount of DNA after amplification (unitless)

Explanation: The formula assumes 100% efficiency where each cycle doubles the amount of DNA.

3. Importance of PCR Product Estimation

Details: Estimating PCR product helps in planning experiments, determining appropriate cycle numbers, and ensuring sufficient product for downstream applications.

4. Using the Calculator

Tips: Enter the initial DNA amount (can be in arbitrary units) and the number of PCR cycles (typically 20-40). The calculator will estimate the final product amount.

5. Frequently Asked Questions (FAQ)

Q1: Is this calculation accurate for real PCR?
A: This assumes 100% efficiency. Real PCR typically has 80-95% efficiency, so actual product may be slightly less.

Q2: What units should I use for initial amount?
A: The calculation is unitless - you can use molecules, ng, or arbitrary units as long as you're consistent.

Q3: Why doesn't the product double exactly each cycle?
A: Factors like reagent limitations, enzyme efficiency, and product reannealing reduce actual efficiency below 100%.

Q4: When does PCR stop being exponential?
A: Typically after 30-40 cycles when reagents become limiting (plateau phase).

Q5: How can I account for PCR efficiency?
A: For efficiency (E) between 0-1, use: Product = Initial × (1+E)^Cycles