I've tried Lipofectamine 2000 followed by G418 selection. Any suggestions?
Thank you
If your results have been poor, it's likely because generic reagents don't work well in embryonic fibroblasts. We used a 3T3-L1 specific reagent (https://altogen.com/product/3t3-l1-transfection-reagent-emb-fibroblast-cells-cl-173/) for siRNA transfection and it gave us good results with greater than 70% efficiency. If you're adding plasmids, then you'll have lower efficiency, but with your selection marker you'll still end up with a population of viable transfected cells.
Hello
See links and discussion on RG , Hope help you
Good Luck
https://www.thermofisher.com/sa/en/home/references/protocols/rnai-epigenetics-and-gene-regulation/rnai-protocol/transfecting-stealth-rnai-or-sirna-into-3t3-l1-cells-using-lipofectamine-2000.html
https://www.researchgate.net/post/How_to_efficiently_transfer_3T3_adipocytes
https://www.lifescience.roche.com/wcsstore/RASCatalogAssetStore/Articles/Protocol_X-tremeGENE%20HP_3T3-L1.pdf
Hi,
I would go for lentiviral transduction if a stable cell line is needed. I have used it in many different cells, including NIH 3T3.
Hope it helps,
Pille
Try HiPerFect transfection reagent. It does give good transfection,. It is a bit more harsh on cells but since you going to select them, it should be ok.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology