1 reply to this topic

[memari]

Hello
I am going to do a stable transduction of fibroblast cells (SCC25). I search in Google and I found a lot of different products and ways which made me confused.
in this link says that I should not use fibroblast cells.
https://docs.google....kmRwMYUC_KXrOGw

SCC25 cells are very tough to transfect buy lipofectamine and similar products.
which product do you recommend?
which transduction is better for SCC25? baculovirus  or lentivirus or ...?

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Babak

Edited by memari, 29 June 2012 - 05:04 AM.

[bob1]

There is no reason to not use primary fibroblasts, other than that primary cells can be difficult to transfect (incidentally I didn't see anything about not transfecting fibroblasts in that link).  If you havn't done transfections before, practice on some cells such as 293 or HeLa with a GFP or other easily detectable plasmid to give you an idea of how to go about it, so as to not waste your primary cells.

If you can afford it electroporators such as the Neon system from Invitrogen work really well, and they have a huge range of optimised conditions for different cell types on their website.

I would go for lentiviral as it is supposed to be very efficient, but I don't have much experience with them myself.