Calculate optimal PCR annealing temperature from primer sequence. Instant Tm calculation using Wallace rule. Free tool with GC content analysis for accurate primer design.
The DNA annealing temperature (Tm) is the optimal temperature for PCR primers to bind specifically to template DNA during amplification. It's calculated using the Wallace rule formula based on the primer's GC content percentage and sequence length. Higher GC content results in higher annealing temperatures because G-C base pairs form three hydrogen bonds versus two for A-T pairs, providing greater thermal stability.
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