...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios...

...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...channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM high ..., 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...