Hello everyone. This question has made my mind busy.
I'm trying to calculate efficiency of my qPCR using extracted DNA of Helicobacter pylori and the results are within accepted range (80-110%).
Since my actual research is on cDNA, do I need to recheck the efficiency using cDNA?
(Considering that bacteria do not contain intron regions and cDNA equals gDNA, I thought maybe the efficiency will not be changed).
Ideally, you should use the same molecule for your standard curve as for your experimental samples.
That being said, most folks will clone their region-of-interest into a plasmid and use digested plasmid DNA for their standard curve because it's possible to calculate exact copy number from plasmid DNA. That means you can calculate exact expression & not just relative expression.
Good luck!
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