Quantified the linearized RLOv plasmids using the Quant-It DNA BR Kit (Invitrogen) according to the manufacturer’s protocol.
Standards (10μL each of 0, 5, 10, 20, & 40ng) were run in triplicate.
Linearized plasmids were quantified in replicates of six.
Quantification was performed in black 96-well plates in the Seeb Lab Victor3 1420 (Perkin Elmer) plate reader. See the spreadsheet linked in the Results below for reading protocol.
Calculations & raw fluorescence readings (Google Sheet): 20151106_quantification_RLOv_linearized_plasmids
Standard Cuve R^2 = 0.999
Best Fit Equation: y = 1174.8215x + 8919.308333333
Will proceed to making qPCR standard curves from the DNA helicase and the head-to-tail linearized plasmids.