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Seeding HeLa cells into 96 well plate for transfection - (Apr/10/2012 )


I would like to seed HeLa cells into a 96 well plates for transfection. I was told that I should use a 80% confluent flask to seed the cells, however, my flask is over confluent and was wondering if it would be OK to use these cells for seeding and then transfecting. i.e. will the efficiency of transfection be effected by this?

p.s. Transfection is done overnight.

Thanks in advance


Potentially yes, the overconfluent cells will have more of a lag phase growth than due to slowing of their growth in very confluent situations, such that this may affect your transfection. Also, do you want to ALWAYS have to do your transfections with over confluent cells - if you start off like that, you should continue so as to minimise the changes between transfections...

Having said that, HeLa cells are very tolerant of a lot of different conditions, so it may not make much difference!


HeLa cell line was derived from a very agressive cancer and they are really tolerant so I don't think it might really affect your results if you need them urgently, but it's a kinda unprofesional approach. If you have time it's better to perform your experiment 100% properly.