Lin Fang Answered Aug 25, 2013
Department of Pediatrics, School of Medicine, Stanford University, USA
Although I have never done such experiment,I'd like to share some of my thought with you. From my understanding of this protocol, it should not affect the experiment fundamentally. Should there is any trivial difference, I prefer plate cell first then treat cells since if the other way (treat cells, then plate them) would involve stress the already stressed (treatment) cells one more time (detach the cells). In addition, for the easiness of operation, it is much easier to plate cell first then the other way around. Since for plate cell first case, you just need to aliquot cells from the same master cell suspension, but if treat cell first, then you need to harvest, count and aliquot from different dish of cells. These are the trivial difference I could think of, but shouldn't change the result.
Hope it may be helpful.