I am working on the expression of protein. I want to know how can we confirm the gene expression of that protein through RT PCR? how can we co-relate the results of RT PCR with the gene expression?
Simple:
Design primers that span exon junction of your Gene of interest. Use these primers with cDNA prepared from cells/tissues of choice and perform RT-PCR and run the products on Agarose Gel. If you see band at the expected size, it indicates your Gene is expressed. This is just qualitative analysis. You should also include a control primer set eg of a housekeeping gene like Gapdh, Actin, TBP etc
Hi Azar,
I agree with Utkarsh Kapoor but take care of all regulations between mRNA production and expression in protein, half-life of the mRNA and its protein, splicing and production of transcrits and proteins....you can of course correlate but don't be surprised by messy results.
choose your primers at the 3' end of the mRNA, where it is the most stable, one across an exon junction and amplicon length about 100-150bp.
fred
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