I am currently working on the quantification of gene copy numbers and
transcripts and got to the point where I need to make my standard curve.
I have already isolated and linearized the plasmids containing the
insert of interest but am (already for a while) wondering for what exact
reason the linearization needs to be done. Unfortunately, I did not find
much information in the literature (at least not in ecological
research). Do the different formations of a plasmid behave differently
in a qPCR reaction? And in how far could the buffer used for the digest
interfere with my qPCR reaction? Do I need to clean my plasmid solution
(and if yes, what's the best way)?
Many thanks in advance!