Home
Back
Q5 Polymerase Melting Temperature Formula:
| From: | To: |
The Q5 polymerase melting temperature (Tm) calculation estimates the temperature at which half of the DNA duplex dissociates into single strands. This formula is specifically optimized for Q5 High-Fidelity DNA Polymerase in PCR applications.
The calculator uses the Q5 polymerase Tm formula:
Where:
Explanation: This formula accounts for the higher thermal stability of GC base pairs (3 hydrogen bonds) compared to AT base pairs (2 hydrogen bonds).
Details: Accurate Tm calculation is crucial for designing effective PCR primers, optimizing annealing temperatures, and ensuring specific amplification of target DNA sequences with Q5 polymerase.
Tips: Enter the exact count of each nucleotide base in your primer sequence. All values must be non-negative integers representing the base composition.
Q1: Why use this specific formula for Q5 polymerase?
A: This formula is optimized for Q5 High-Fidelity DNA Polymerase and provides accurate Tm predictions for primer design with this specific enzyme.
Q2: What is the typical Tm range for Q5 PCR primers?
A: Optimal primer Tm for Q5 polymerase is typically between 55-72°C, with annealing temperatures usually 3-5°C below the calculated Tm.
Q3: How does this differ from other Tm calculation methods?
A: This is a simplified nearest-neighbor method specific to Q5 polymerase. Other methods may use more complex algorithms or different coefficients.
Q4: Can I use this for degenerate primers?
A: For degenerate primers, calculate Tm for each possible sequence and use the average, or design with the most stable configuration.
Q5: What if my primers have modified bases?
A: This calculator is designed for standard DNA bases. Modified bases may require specialized Tm calculation methods.