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SOLVED: A widely used method for calculating the annealing temperature for a primer used in PCR is 5 degrees below the Tm (ºC), which is computed by this equation 81.5 + 0.41 (%
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Versatility of different melting temperature (Tm) calculator software for robust PCR and real-time PCR oligonucleotide design: A practical guide | Mohammad Javad Najafpanah - Academia.edu