https://pages.htgaa.org/2026a/prashant-shah/homework/week-06-hw-genetic-circuits-part-i/index.htmlAssignment: DNA Assembly What are some components in the Phusion High-Fidelity PCR Master Mix and what is their purpose? Phusion HF Master Mix contains several key components. The centerpiece is Phusion DNA Polymerase, a high-fidelity enzyme that includes a 3’ ti 5’ proofreading (exonuclease) activity, meaning it can detect and correct misincorporated bases during synthesis — this gives it roughly 50x lower error rates than standard Taq polymerase. The mix also contains dNTPs (dATP, dCTP, dGTP, dTTP), which are the nucleotide building blocks that the polymerase incorporates into the new strand. A reaction buffer (HF buffer) maintains the optimal pH and ionic conditions, and includes MgCl₂, a cofactor required for polymerase activity and primer-template stability. Finally, it contains stabilizers and enhancers that improve yield and specificity. Using a 2X master mix simply means all these components are pre-mixed at twice the working concentration, so you just add your template, primers, and water. To Summarize, DNA polymerase: extends primers to copy the template DNA. • dNTPs: raw materials for making the new DNA strands. • HF buffer: supports enzyme activity and specificity. • Mg²⁺: required for polymerase catalysis. • Water: brings the reaction to final volume. • Template + primers are added separately in the protocol. Hugoen