Fig. 3.

Workflow for determining linear plasmid copy number and dilution. Calculate the copy number concentration of the linearized plasmid and the volume needed for a dilution of 2E+9 copies per μL (1E+10 copies per 5 μL), based on the known length of the plasmid, “Plasmid Length (bp),” and the mass concentration of the linearized plasmid as determined in Subheading 3.2, steps 6 and 7, “measured mass concentration (g).” Choose pipettes that maximize the accuracy and precision of reagent delivery. *See Subheading 3.2, steps 6 and 7 and Notes 19 and 20. **The final concentration of the stock linear plasmid dilution may be adjusted according to the linear plasmid yields obtained. For example, this value may be reduced from 2E+09 to 2E+08 copies/μL to create a stock of 1E+09 copies/5 μL, rather than 1E+10 copies/ 5 μL, if the yield of linear plasmid is low. ***See Subheading 2 for exact formulation