3 replies to this topic

[xyz74]

Hello,

I am doing transfections for the first time and I have a question about lipofectamine 2000.

It says to change media after 4-6 hours. I have a total of 600ul solution in each well of my plate. So should I just aspirate the entire solution out and then replace it will my fresh media? Or should I add more media to the wells and not remove anything from the well?

Thanks.

[genehunter]

Just do the  remove and replace.

[scolix]

Sometimes I change the media and some times I donot change the media after lipofectamine transfection. Both conditions work for me.

[SS17]

ideally you should replace with fresh media and not do the top up thing, it would depend on your cell type, on some sensitive cells leaving the lipid reagent would make the cells unhappy and plus  you wouldnt want tiny lipid particles floating about in your culture media which might scare the life out of you if you end up thinking that its some sort of contamination!