Testing primers on 'unknown' tissue - (Jul/15/2014 )
I want to perform RT PCR on a fibrous-like tissue.
I don't know exactly whats in it (cells/matrix) but histology gave me an idea.
Therefore I designed some primers for identification of fibroblasts-macrophages-osteoblast-endothelial cells.
I just want to know if these genes are expressed in my tissue (and how this differs between donors)
However, how do I test these primers for my tissue?
Do I just take cDNA from one of my donors to test all primers (problem is I don't know if they are expressed) or do I have to test these primers on for example fibroblasts, macrophages, osteoblasts seperatly?
If you want to test that the primers work, then you could use either a cell line that expresses the gene(s) you are interested in or something like a plasmid containing gene of interest.