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Question of transfection reagent

Can FreeStyle MAX reagent be used for transfecting adherent cells?

Yes, the reagent can be used for adherent cells, however the protocol is an optimized protocol for CHO and 293 cells in suspension culture. If you want to transfect adherent cells please use Lipofectamine 2000 transfection reagent. Please use this link for the transfection protocol with Lipofectamine 2000.

http://www.invitrogen.com/content/sfs/manuals/lipofectamine2000_man.pdf

 
How does FreeStyle MAX reagent compare to Lipofectamine 2000 and 293fectin?

Lipofectamine 2000 is designed for adherent cells. FreeStyle 293fectin is designed for Hek 293 suspension cells. For transfecting 293 suspension cells, FreeStyle MAX reagent yields transfection efficiency comparable to Lipofectamine 2000 and 293fectin. However, Lipofectamine 2000 and 293fectin reagents do not work well for CHO suspension cells.

 
Why do I see precipitate on the cells after transfection?

A small granular-like precipitate may be detected microscopically on the cells after transfection using cationic lipid. This is normal. The presence or absence of this precipitate is not indicative of the transfection efficiency.



The precipitate can be caused by presence of excess EDTA or cationic lipid. Use DNA in water or, if in TE, use EDTA concentrations of <0.3 mM in the diluted DNA. Ensure concentrations of cationic lipid reagents do not exceed recommended amounts in complex formation.

 
What is the recommended ratio of DNA and transfection reagent when transfecting

We recommend 1:1 ratio of DNA and lipid. For example, 1.25 ug DNA and 1.25 ul of the reagent for one ml of suspension cells.

 
What is the recommended ratio of DNA and transfection reagent when transfecting

In general, the transfection efficiency for FreeStyleMAX reagent is 65% when used with suspension CHO and 293 cells.