AAV Production in HEK293 Cells
Type
Protocol
...of each plasmid needed to have a 1:1:1 molar ratio with 2 mg total DNA per CS5 Plasmid Plasmid Size (bp...conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene ...molar ratio of each plasmid: 2000 μg / 24,961 bp = 0.08 μg/bp Therefore, for each plasmid we need: Sample ...Volume of DNA (μL) RepCap 7,265 1.00 727.6 pHelper 11,854 1.00 1,185 Transfer Plasmid 5,842 1.00 584.2 ... the total number of base pairs from all three plasmids, we can determine the total μg/bp we need to achieve...Needed: 949.8 μg / 1.0 μg/μL = 949.8 μL Transfer Plasmid: 0.08 μg/bp × 5,842 bp = 468.1 μg Volume Needed... 2% HI-FBS into a sterile 1 L bottle. Add each plasmid DNA into the bottle containing the Opti-MEM. Mix...