1. What is PCR Amplification?

PCR (Polymerase Chain Reaction) amplification is a method to exponentially amplify DNA sequences. The amplification factor represents how many copies of the target DNA are generated after a given number of cycles.

2. How Does the Calculator Work?

The calculator uses the PCR amplification equation:

Where:

• \( 2 \) — Represents the doubling of DNA in each cycle (assuming 100% efficiency)
• \( \text{cycles} \) — Number of PCR cycles performed

Explanation: Each PCR cycle theoretically doubles the amount of DNA, leading to exponential growth in the number of copies.

3. Importance of PCR Amplification

Details: Understanding amplification is crucial for determining the sensitivity of PCR assays, optimizing cycle numbers, and interpreting results in applications like genetic testing, forensics, and research.

4. Using the Calculator

Tips: Enter the number of PCR cycles (typically between 20-40 for most applications). The calculator will show the theoretical amplification factor.

5. Frequently Asked Questions (FAQ)

Q1: Why is the amplification 2^n?
A: This assumes perfect doubling efficiency (100%). In practice, efficiency might be slightly less due to various factors.

Q2: What's a typical number of PCR cycles?
A: Most protocols use 25-35 cycles. Too few cycles may not produce enough product, while too many can lead to nonspecific amplification.

Q3: How does efficiency affect amplification?
A: If efficiency is less than 100%, the formula becomes (1 + efficiency)^cycles. 100% efficiency means doubling (1+1=2).

Q4: Why does amplification plateau?
A: In later cycles, reagents become limiting and polymerase activity may decrease, causing the reaction to plateau.

Q5: Can I calculate starting DNA from amplification?
A: If you know the final amount and amplification factor, starting amount = final amount / amplification factor.