Tm Calculator Method Review: 50 Primers
Use this report when you need to explain why NEB, IDT, Primer3, or a legacy calculator can return different Tm values for the same primer. This review compares 5 Tm calculation methods across 50 primer sequences spanning 7 categories (standard PCR, GC-rich, AT-rich, short, long, qPCR probes, and CRISPR guides) under the same standard conditions (50 mM Na⁺, 250 nM oligo).
This report supports the Tm Calculator with method validation data. Use it to compare calculation assumptions, then calculate your own sequence in the Tm Calculator.
Dataset Basis and Limits
The benchmark uses representative synthetic primer sequences selected to cover common design categories: standard PCR, GC-rich, AT-rich, short, long, qPCR probe-like, and CRISPR guide-like sequences. It compares formula behavior under fixed assumptions; it is not a wet-lab validation set and does not claim PCR success, synthesis quality, or vendor equivalence.
Calculating method comparison...