I have prepared primary cortical neurons from mice embryos and would like to express plasmids via transient transfection. Some people note that lipofectamine can be toxic and interfere with downstream experiments viz. patch clamp or functional imaging. Does anyone know a better alternative? How good is Clontech's CalPhos reagent? Thanks in advance!
Hi Madhuvanthi, you have to use either gene gun method or viral infecction. The last one it is more time-consuming because necessary to make virus but it is more efficient.than the first. With the first one you have to make high-density cell culture of cortical neurons.
Hello!
Have you already though using mRNA instead of plasmid DNA?
I would strongly recommend to read this PhD thesis, in particurlar the last chapter which is about transfecting neurons in vivo.
https://smartech.gatech.edu/handle/1853/58715
If you need more info about the Viromer RED, please ask me.
Bests, Sandra