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12 well transfection - (Dec/12/2006 )

goodmorning all
i really appreciate if u can help me
i used to do transfection of 293 cells in 6 wells plate and it was ok, now i am using 12 well plate and i find difficulties in seeding the cells evenly in each well, i mean i tried several options but still i found my cells gathering in the centre of the well next day.
thankx for any comments.


for instance, you may try to tap the plate with your hands to enhance homogenization.
Check also with very few of medium if the whole plate is covered or if just the center is overed, which may mean that the plastic is not flat.


Try to increase the volume of medium to 2 ml/well, sse if that makes any difference. Avoid excessive mixing, that causes so called "vortext effect" and your cells end up in the center.


Tilt ur plate to one side (front and back) till the media is abt to half way thru the well and then do it the other side. After abt 10 times, try to go side ways abt 10 times, then again 5 times back and front. After this look atthe plate under microscope to get an idea of how the cells have distributed.

i do this for 24 well plate seeding and get an even plating of cells.


I use Scolix's method too, it really works.


thankx everybody
i will try it on monday.