https://pages.htgaa.org/2026a/zarias-arman-aelius-saadatkhah/homework/week-06-hw-genetic-circuits-part-i/index.htmlPart A — DNA Assembly Questions Q1: What is in a Phusion PCR master mix and what does each component do? A standard Phusion High-Fidelity PCR master mix contains the following components: Component Role Phusion Hot-Start DNA Polymerase High-fidelity thermostable polymerase with a 3’→5’ proofreading exonuclease; error rate ~4.4 × 10⁻⁷ per bp per cycle (50× lower than Taq) dNTPs (dATP, dCTP, dGTP, dTTP) Nucleotide substrates for strand synthesis Mg²⁺ (MgCl₂, 1.5–3.0 mM) Essential cofactor for polymerase activity; stabilises the primer–template duplex 5× HF Buffer (KCl + Tris-HCl pH 8.8) Maintains optimal pH and ionic strength; HF formulation includes a proprietary enhancer that increases specificity DMSO (optional, 0–3%) Denaturant for GC-rich or secondary-structure-prone templates Primers (user-added, 0.5–1 µM each) Define amplicon boundaries; anneal to template strands Template DNA (user-added, 1–50 ng) Source of target sequence Nuclease-free H₂O Brings reaction to volume The hot-start formulation keeps polymerase inactive below ~60°C, preventing non-specific extension during setup and eliminating the need for a manual hot start.Hugoen