Transfection

•2x105 cells were seeded in a 6 well plate and left overnight in DMEM F12 +10%FBS at 37ºC and 5% CO2

•Solutions A and B were prepared as follows                              

- A= 2µg DNA + OptiMEM to make up the volume to 200µl

-B= 8µl of cellfectinII + 192µl of OptiMEM

•Incubated for 5 min at RT

•A and B were mixed, tapped well and incubated for 30 min at RT

•Cells were washed with DPBS twice

•600µl of OptiMEM was added to 400µl of A+B mix and added to each well

•Incubated for 6 hrs at 37ºC and 5%CO2

•Mix was removed  and cells were washed twice with DPBS

•Fresh DMEM F12+ 10% FBS was added and incubated at 37ºC and 5% CO2 for 48hrs