I'm running a co-culture experiment with BV2 and N2a cells. I'm trying to understand how LPS treatment of BV2 can influence expression of BDNF and NGF in N2a. The problem is that I can't detect these genes in qPCR (Ct > 38).
Have you used these primers before in your lab with good results? in case they are new, have you tried with different primers for each target?
Cristian Pérez-Fernández Yes I did that ! I really think that the mRNA are not detectable in the cells and I don't really know why.
But your HK is expressing at Ct17, no? so your mRNA seems to be OK (I guess you checked integrity and purity). The issue here is about your GOIs imo. I also guess that, when you say that these genes expressed at>Ct38, you are refering to control samples (cells without LPS exposure/exposed to vehicle), isn't it?
Cristian Pérez-Fernández Yes I checked for the RNA. Yes I refered to control samples. You might be right these genes shouldn't be expressed in these cell lines. Thank you
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