I need to transfect several novel viruses in S2 cells via electroporation and I am looking for a protocol for that. Any suggestion?
Thanks
I have never heard of electroporation for insect cells. We usually use transfection to insert genes in cell's genome. You need a transfection agent like cellfectin or lipofectamin. They come with manuals which have a protocol for different cell lines.
It seems that both reagent-based and electroporation transfection methods have been applied for S2 cells (Attached is a reference paper). I already tried lipofectamin to transfect viruses with small genomes which I was bale to clone them and it worked. However, now I have two other viruses with big genomes (11 and 27 kb) that I have many problems to clone them because of toxicity in E. coli. So I have decided to try to infect cells with filed collected infected insects as an alternative approach. I read that people used this technique for Falviviruses and it worked. The only thing that I don`t know is that how they infected cells with that filtered solution, probably electroporation?
Other point is that I do not know if these two viruses can replicate in S2 cells as they have been identified in a completely different insect which there is no any available cell line for that.
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