Dear Daniel,

We always use LipoGeneTM Transfection Reagent, it shows very high transfection efficiency. You can find detailed information in the following website: https://www.genemedi.net/i/lipogene-transfection-reagent

Hope this may help you.

Lentiviral vectors from Addgene. www.addgene.org

Lipofectamine LTX/Plus works well in NIH/3T3 cells (65% GFP transfection efficiency), it will give you higher transfection efficiency in your MEF cells. If you use 24-well plate format to do your experiment, the transfection protocol will be DNA/well : 0.5ug+ 0.5ul Plus in 25ul Opti-MEM; Lipofectamine LTX /well: 1.75ul in 25ul Opti-MEM ; Combine these two solutions, 5min RT incubation, then complex to cells; Post-transfection 48hrs to analyze your experiment.

Please take a look the detail protocol at: http://tools.invitrogen.com/content/sfs/manuals/lipofectamineLTX_and_PLUS_man.pdf

Based on the protocol, you can scale up and down your experiment.

If you are doing siRNA transfection , you might use Lipofectamine RNAiMax as your transfection reagent. The detail protocol will be at:

http://tools.invitrogen.com/content/sfs/manuals/lipofectamineRNAiMAX_man.pdf

Goodluck!

I agree with all. If you dont have a lentiviral system, lipofectamine is the best option.

Thanks for your answers!

I will try Lipofectamine with my MEF cells. If this won't work that well I will check if transduction is possible method in the lab.

I used to use Dharmafect 3 with many cervical cancer cell lines for siRNA transfection. Lipid concentration was 0.05%, I found this concentration optimal with very low cytotoxicity and yet not diluted. Lipofectamine 2000 was the second best option.

Good luck :)

Hi, lipofectamine is only for transient transfection, right? If I want to do stable transfection, is lentivirus the only option? Or is it the best option?

For MEF, I recommend either DreamFect or Magnetofectamine which is a magnetic nanoparticles formulation combined with lipofectamine to increase the efficiency. if you want publications showing that these reagent works well on MEF let me know [email protected]

Thank you for this reccomendation. I will have a look for them.

I tried quite a lot of different reagents and I got the best results using such magnetic nanoparticles (NeuroMag, Nanotherics) or Viromer RED (Lipocalyx).

Both work quite well for the transfection of MEF.

Hi Daniel,

Yes I agree NeuroMag is also pretty good for those cells.

Dear Daniel, you can please follow the links below to obtain certain valuable informations:-

1. http://signagen.com/index.php?main_page=product_info&products_id=768

2. http://ezbiosystems.com/view2.asp?d_id=344

3. https://lifescience.roche.com/wcsstore/RASCatalogAssetStore/Articles/Protocol_X-tremeGENE%20HP_MEF.pdf

Further you can also go though the document attached herewith.

Lipofectamine 3000 is meant to be more efficient than 2000. Follow this link to check it out. http://www.lifetechnologies.com/uk/en/home/brands/product-brand/lipofectamine/lipofectamine-3000.html#resources

Here's a kit for MEF cells (https://altogen.com/product/mef-transfection-reagent-mouse-fibroblast-cells/). It has the transfection reagent and buffers to improve efficiency if you need it. Be sure you know what you're trying to achieve - the question is a bit vague, but if you're doing siRNA transfection high efficiencies should be sufficiently easy to reach.

Dear Daniel,

We always use LipoGeneTM Transfection Reagent, it shows very high transfection efficiency. You can find detailed information in the following website: https://www.genemedi.net/i/lipogene-transfection-reagent

Hope this may help you.