I am planning to transfect my mammalian cells with plasmid DNA but for various reasons, I only have a small amount (about 6 ul of 100ng/ul). I had planned to try a transfection on a 6-well plate using 2ul of DNA and 6ul of Turbofect 8.0 (keeping the 3:1 ratio), however this means there will only be approx 200ng of DNA per well when I transfect, obviously the standard is usually 1ug/ul but I do not have this quantity. Can you tell me please if this has been tried before to your knowledge and if so, has it been successful? Any advice you could offer would be much appreciated, thank you!
Yes, in general you can use smaller quantities of DNA, but don't expect to achieve high transfection efficiency. I've used manufacturer protocols for 24 well plates (https://altogen.com/product/altofect-transfection-reagent/) and I would say that if you don't have that much DNA, just don't use that many cells, and you'll still have the same amount of transfected cells at the end.
I believe it is possible but it may rely on several parameters including but not limited to: the quality of the vector design and purification, the goal of transfection, target cell type, the quality and brand of the lipid-based transfection method, etc.
Great thanks. I'm going to try using 24 well plates and 300ng which is the most DNA I can spare. Protocol recommends 250-1000ng per transfection so fingers crossed this will work. Thank you!
Yes, in general you can use smaller quantities of DNA, but don't expect to achieve high transfection efficiency. I've used manufacturer protocols for 24 well plates (https://altogen.com/product/altofect-transfection-reagent/) and I would say that if you don't have that much DNA, just don't use that many cells, and you'll still have the same amount of transfected cells at the end.
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