significance of confluence in transfection efficiency - (May/05/2008 )
hi all,
i'm new to transfection and am facing problems from the start. i had plated vero cells in 24 well plates around 12 hours back, and they are already at 100 percent confluence. in this condition, is there any point in doing transfection? i am using lipofectamine, and invitrogen recommends 90 - 95 percent confluence at the time of transfection. i assume this will definitely affect the efficiency. so should i go ahead with the transfection, or just wait till a new plate with reduced number of cells is ready?
do help me out. thanks anyways.
I would prefer to plate new cells. Your point is good to optimize the cell number for the experiment. This is also important for the reproducibilty of your experiment.
Please correct me if I am wrong but I think the high confluency recommended for transfection with Lipofectamine is due to cell death after transfection.
Confluent cells are much less transfectable than 80-90%.
i'm new to transfection and am facing problems from the start. i had plated vero cells in 24 well plates around 12 hours back, and they are already at 100 percent confluence. in this condition, is there any point in doing transfection? i am using lipofectamine, and invitrogen recommends 90 - 95 percent confluence at the time of transfection. i assume this will definitely affect the efficiency. so should i go ahead with the transfection, or just wait till a new plate with reduced number of cells is ready?
do help me out. thanks anyways.
try to transfect cells between 40 and 80% confluency which means sthat they are in a well proliferating state