Reporter assay selection with LipoD293 DNA transfection reagent

Use lipoD293 DNA transfection reagent for transfection of multiple mammalian cell lines (such as Hela and PC3) with luciferase reporters, you will get much higher luciferase readings than other transfected reagents used on our lab for luciferase reading, but observe little cell death in transfected cells in our condition!

Use unsupplemented RPMI1640 / DMEM instead of Opti-MEM medium to dilute both lipoD293 and DNA (luciferase reporters), you will get further higher luciferase readings (one-fold increase) and save more money !

Several key points in the protocol:
1) Use the same type of culture medium (but unsupplemented) for diluting lipoD293 reagent and luciferase reporter DNA( for example, if the cells are cultured in RPMI1640-based medium, then use unsupplemented RPMI1640 as dilution medium);
3) No need to change medium either before or after transfection;
4) Add diluted lipoD293 reagent to diluted DNA and then incubate the mixture at room temperature for 15~20 minutes;
5) Make lysates for luciferase assay 16~48hrs post transfection