I need to transient transfect HEK293 cells with a plasmid containing a luciferase tag and a plamid containing a histamine receptor. I have never done this before so I am looking for a straight forward and detailed protocol, but seems that in many paper nobody mention any.
Can someone help me? We Lipofectamine LTX in the lab and we can do electroporation.
Hi, I usually transfect even 3 plasmids together. Just split the total amount of DNA that you have in your protocol by the numbers of plasmids.
Suppose:
3 ug of total DNA
- 1.5 ug of luciferase plasmid
- 1.5 ug of his plasmid
Then add the amount of transfection reagent for 3 ug of DNA following your protocol.
a little side note, for us, transfection with linear PEI instead works pretty well for HEK cells as an alternative to lipofectamine + electroporation. PEI is not very expensive and it allows you to transfect larger amounts of cells.
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