stimulation experiments - How do I go about doing time course and dose experiments? (Aug/08/2007 )

I want to stimulate my mammalian cells with bacteria and then extract the RNA and use qPCR to look at the effect on expression of certain cytokines and chemokines. I need to first establish how long I need to stimulate my cells for and what multiplicity of infection I should use. I don't know how to go about this. I dunno which experiment is the right one to do first or if I should do them both together. I am very confused and don't know how to progress. Any advice? Thanks!

hey LittleMiss. are you still working with S aureus and adherent epithelial cells? if so, I can tell you exactly how to do this - it was what I did at my last job and I can even send you to the papers for the protocol let me know - we used S aureus and NHEK cells, and we looked at both cytokine and defensin expression using qRT-PCR, among other things

us read-headed cats need to stick together; you, me, and fred

hey LittleMiss. are you still working with S aureus and adherent epithelial cells? if so, I can tell you exactly how to do this - it was what I did at my last job and I can even send you to the papers for the protocol let me know - we used S aureus and NHEK cells, and we looked at both cytokine and defensin expression using qRT-PCR, among other things

us read-headed cats need to stick together; you, me, and fred

hey Littlemiss

I'm just coming off vacation and am away from the lab till Monday; I've been gone a week. this is why I have not yet rushed you the protocol

I'll pm it to you on Monday (my protocol-bible lives at the lab)

take it easy
A