...1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios... 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume 500 μL *Plasmid concentrations and ... each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid DNA purification...endotoxin removal step. For high quality plasmid DNA, the plasmids should also be propagated in an endonuclease... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...~20 h. Prepare a mixture of the 3 transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish...is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1-2 ...

...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ...protocol to transfect mammalian cells with your plasmid of interest. Protocols...polyethylenimine. Transfections allow for transient expression of a gene of interest in a target cell line and..., the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable....is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 ...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X 293T cells were ...