Table 3.

DNA amounts for lipid transfection, based on prime editor system

PE System	Amounts of Transfection Components
PE2	200 ng PEmax plasmid 66 ng epegRNA plasmid
PE3/PE3b	200 ng PEmax plasmid 66 ng epegRNA plasmid 22 ng nicking sgRNA plasmid
PE4	200 ng PEmax plasmid 66 ng epegRNA plasmid 100 ng MLHIdn plasmid
PE5/PE5b	200 ng PEmax plasmid 66 ng epegRNA plasmid 22 ng nicking sgRNA plasmid 100 ng MLHIdn plasmid
TwinPE	200 ng PEmax plasmid 33 ng epegRNA 1 plasmid 33 ng epegRNA 2 plasmid
TwinPE + recombinase single-transfection targeted donor integration *Optimized for 48-well plate	500 ng PEmax plasmid 50 ng epegRNA 1 plasmid 50 ng epegRNA 2 plasmid 200 ng Bxbl plasmid 200 ng recombination donor plasmid

All of these amounts, except for those associated with single-transfection integration, have been optimized for 96-well plate transfections of HEK293T cells using 0.5 μl per well of Lipofectamine 2000.The single-transfection integration amounts have been optimized for 48-well plate transfections of HEK293T cells using 1 μl per well of Lipofectamine 2000.