https://pages.htgaa.org/2026a/alvaro-pacheco-concha-buonamici/homework/week-6--genetic-circuits-part-i-assembly-technologies/index.htmlPhusion Master Mix Components This mix contains a high-fidelity polymerase with proofreading activity (3 to 5 exonuclease) to minimize sequence errors. It also includes dNTPs as DNA building blocks, an optimized buffer with salts like magnesium chloride that act as enzymatic cofactors, and stabilizers to maintain the pH and ionic strength required for the reaction. Factors Determining Annealing Temperature The annealing temperature primarily depends on the melting temperature (T_m) of the primers, which is influenced by sequence length and GC content. Other external factors include the concentration of salts (monovalent and divalent) in the PCR buffer and the concentration of the primers themselves in the mixture.Hugoen