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[bigbadbob]

Apologies for the basic question but i want to use a real-time PCR assay using absolute quantification to quantify DNA quantities in patient samples.
I have cloned my PCR amplicon into a plasmid and confirmed its identity through sequencing.
I then prepped this plasmid up and quantified on a nano-drop to give the exact number of copies of my amplicon.
When using this plasmid as a standard dilution series in my assay to calculate how much DNA there is in my samples do I need to also extract my plasmid first before use in constructing my curve.

Thanks