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Hello Folks,
Just a quick one that I am hoping someone can answer:
I am interested in quantifying the number of bacteria in a population using qPCR. I have primers to a gene that is present once per genome (DNA gyrB) and amplifies up a 150 bp fragment. I will make a standard curve using purified and quantified PCR amplicons for this assay.
My question is that I have read papers in which people go to, what I consider to be great lengths to obtain template for their calibration curve by cloning the fragment into a plasmid and then extracting and purifying it this way. Why do that when you could just PCR up your product, verify via a gel and purify and use it that way? It's making me question just using PCR product for this.
Thank you
One.
