1. What is PCR Product Calculation?

PCR (Polymerase Chain Reaction) product calculation determines the length of the amplified DNA fragment based on the primer binding positions. This is essential for experimental design and verification of PCR results.

2. How Does the Calculator Work?

The calculator uses the simple formula:

Where:

• Start Position — The 5' end nucleotide position of the forward primer
• End Position — The 3' end nucleotide position of the reverse primer

Explanation: The +1 accounts for inclusive counting of both start and end positions in the product length.

3. Importance of PCR Product Calculation

Details: Accurate product length prediction helps in gel electrophoresis analysis, primer design validation, and ensuring the amplified fragment matches the expected size for downstream applications.

4. Using the Calculator

Tips: Enter the start and end positions in base pairs (bp). Both values must be positive integers, and the end position must be greater than or equal to the start position.

5. Frequently Asked Questions (FAQ)

Q1: Why add 1 to the position difference?
A: The +1 accounts for the inclusive nature of position counting. If you amplify from position 100 to 200, the product includes both positions, resulting in 101 bp.

Q2: What if my primers overlap?
A: Primers should not overlap in standard PCR. The end position must be greater than the start position for a valid product.

Q3: Does this account for primer lengths?
A: No, this calculation assumes you're entering the binding positions of the primers. The actual product length is determined by the distance between primer binding sites.

Q4: Can I use this for qPCR product calculation?
A: Yes, the same calculation applies to qPCR products as they follow the same amplification principles.

Q5: What are typical PCR product sizes?
A: Most PCR products range from 100-1000 bp, though specialized techniques can amplify larger fragments up to several kilobases.