We are looking into the response induced by two plasmids (a backbone and a construction) and we wondered whether immune response is related to the quantity of mass (in bp) or related to the number of plasmids inserted into the cell (regardless their size).
Hi there,
As you want to compare the responses, the applied dose has to be the same in terms of number of molecules. So constant molar dose is the right way to proceed (ie. ratio mass injected/plasmid size in bp has to be the same for the 2 plasmids).
Thank you Dominique Liger.
We usually transfect the same mass of plasmid (in ug) when comparing treatment vs. control (empty vector). But, in this case and according to your answer, we should calculate with size correction as to keep constant the number of molecules transfected.
Being the backbone smaller in size, the cells receiving this vector will capture less mass of exogenous DNA, right?
For example, if I have a backbone of 4kbp and the construct bearing 8kbp, keeping the no. of molecules constant means that cells receiving the construct would be offered (e.g.) 3ug of plasmid meanwhile cells receiving the backbone would be offered 6ug of plasmid. I wondered if this distinction in DNA captured would influence over the DNA sensing and immune response.
Again, thank you.
Santiago
Hi again,
Actually 3µg for the empty plasmid and 6µg for the construct.
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