Data Source and Methodology
All calculations are based on the nearest-neighbor thermodynamic models, as outlined in the latest scientific literature.
The Formula Explained
Tm = 64.9 + 41 * (G+C-16.4) / (A+T+G+C)
Glossary of Terms
- Primer Sequence: The sequence of nucleotides in the DNA primer.
- Primer Concentration: The molar concentration of the primer in the solution.
- Salt Concentration: The molar concentration of salt (usually NaCl) in the solution.
Frequently Asked Questions (FAQ)
What is the melting temperature (Tm)?
The melting temperature (Tm) is the temperature at which half of the DNA strands are in the double-helix state and half are in the 'melted' single-strand state.
How do I calculate Tm manually?
You can calculate Tm using the formula: Tm = 64.9 + 41 * (G+C-16.4) / (A+T+G+C), where A, T, G, and C are the respective nucleotide counts in the primer sequence.
Why is Tm important?
Tm is critical for optimizing PCR conditions and ensuring the specificity and efficiency of the reaction.
What affects Tm?
Tm is affected by primer length, GC content, primer concentration, and salt concentration in the solution.