Transfection Protocol for HEK293 using GenCarrier-1? - HELP (Sep/12/2005 )
I recently purchased the GenCarrier-1 transfection reagent because I've read so many good things about it's efficiency. I am still at the stage of testing what ratios of DNA to GenCarrier-1 to use as indicated in the protocol, and I haven't been able to see any fluorescence whatsoever. I am currenly just using a control GFP vector and HEK293 cells to find optimal conditions. I've tested 1ug/100uL of DNA to 0.5uL, 1uL, 2uL, 4uL and 6uL of GenCarrier. I've incubated for as short as 5 hours and as long as 18 hours, and nothing has worked. It's getting a little frustrating, and I'm hoping someone can help me out.
Just let me know if I understood, you transfected 293 cells with this supposed great transfection reagent and you saw nothing. What do you try to transfect? GFP? What cells do you plan to transfect with it? 
We don't have any problem with gencarrier-1 on 293 cells. Be sure to use PBS-free, antibiotic-free and serum-free solution during DNA/liposome formation. And don't freeze the reagent. Then you should be fine.
I also used gencarrier-1 on 293, COS, 3T3, CHO-k1 and HeLa cells with very consistent and satisfactory results. One thing I can think of is your 293 cells, they are the easiest and worst cells to work with sometimes (detached, very sensitive, fragile...). Thaw a new vial of 293 and do the transfection again.
In our hands, gencarrier-1 works much better than lipo2000, fugene6, jetpei, genejuice....in efficiency, toxicity and cost. The cell lines we've transefcted in our lab include C2C12, HUVEC, 293, COS and HCT-8.
Your 293 cells may be aged.
Sorry but I can't figure out why you want to use this expensive transfection reagent to transfect 293 cells? Why don't you use calcium phosphate (eventually with chloroquine)?? Much cheaper...
I am sick and tired of this "gencarrier" marketing junk. What I absolutely hate is when the same person posts to a thread 4 or 5 times under different names. Screw this marketing crap. Most publications are using Lipofectamine 2000 reagent. And you know why? Because its BETTER!
Yes, much cheaper and nearly 100% transfection efficiency, as measured by GFP expression
I am sick and tired of this "gencarrier" marketing junk. What I absolutely hate is when the same person posts to a thread 4 or 5 times under different names. Screw this marketing crap. Most publications are using Lipofectamine 2000 reagent. And you know why? Because its BETTER!
[/quote]
I apologize for this rude, insulting and baseless reply and would like to apologize to j0liefllle, postdoc2130, joy_maf, bonita and to this board.
I am sick and tired of this "gencarrier" marketing junk. What I absolutely hate is when the same person posts to a thread 4 or 5 times under different names. Screw this marketing crap. Most publications are using Lipofectamine 2000 reagent. And you know why? Because its BETTER!
Yeah, right. Ha-Ha-Ha .... Check out the difference between me "tom9217" and NOT ME "tom9217." Another piece of shti. Don't change my words please, what I said was:
"I am sick and tired of this "gencarrier" marketing junk. What I absolutely hate is when the same person posts to a thread 4 or 5 times under different names. Screw this marketing crap. Most publications are using Lipofectamine 2000 reagent. And you know why? Because its BETTER!"