Annealing Temperature on Michael’s Domainhttps://jeltsch.org/en/tags/annealing-temperature/Recent content in Annealing Temperature on Michael’s DomainHugoen-usCopyright © 2002 - 2026 Michael Jeltsch.Fri, 24 Jul 2026 00:18:18 +0300Formula to Calculate the Annealing Temperature of Oligonucleotides for PCRhttps://jeltsch.org/en/annealing_temperature/Wed, 30 Aug 2006 00:00:00 +0000https://jeltsch.org/en/annealing_temperature/<p>The thumbrule for calculating the annealing temperature for a PCR primer isTm (°C) = 81.5 + 0.41(%GC) - (675/N) where %GC is the percentage of G and C nucleotides in the oligo and N is the length of the oligo given in nucleotides.</p>