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Testing primers on 'unknown' tissue


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#1 Maemo

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Posted 15 July 2014 - 01:58 AM

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?

 

 



#2 bob1

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Posted 15 July 2014 - 03:04 AM

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.






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