Stratagene Reference RNA - (Jun/12/2006 )
I'm getting a bit bogged down with this stuff now. I'm new to PCR, and use Applied Biosystems pre-made gene expression kits for real time PCR. However, i need to set up a relative efficiency plot for each of my probes, and therefore have ordered in the Stratagene Reference RNA. Now, I'm confused. It suggests that you do the cDNA step and the Real Time in one go, but I haven't got anyone here to help set up the ABI7700 as they seem to suggest.
So...should I use our method here for cDNA sysnthesis form a range of dilutions of the Reference RNA? Then use this template on the ABI machine? Has anyone actually got a protocol for the stratagene ref rna?
Sorry, seems like a daft question, but there's no one here who's done any kind of PCR!
I have not used commercial RNA as a reference, but I think that probably you need to treat it exactly as you do your test RNA samples or it won't be a good reference...it's purpose is as a positive control, right?