Hi all,
I did a mock transfection using pGL3 vectors with different promoter inserts (plasmids range from 10 to 14kb). However, I have gotten unexpectedly low luminescence ratios (firefly luc / renilla luc) and I was wondering if it would be logically sound to transfect mols of DNA instead of a certain weight for all of them (eg. 200ng per well). Thoughts?
Thanks!
Hi Andrew Lin ,
Short answer: Yes, especially if you want to compare these, it makes a lot of sense to go by moles. Still, transfection may decrease with plasmid size as well...
Hope that helps!
Thomas Mühlenberg have you tried doing this before? If so, do you have any guidelines for a lipofectamine:DNA ratio?
No, I haven't. But it is logically sound.
I would just use ~40% more µg of the 14kb plasmid, compared to the 10kb version....
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