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Position: Discussion forums > Bioscience biotechniques discussion forums > Molecular biology troubleshooting forum
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siRNA transfection
Posted by: Caro (IP Hidden, Unregistered user, )
Date: January 23, 2005 03:43PM
Hi there
Has anyone tested different transfection reagents for transfection siRNA, like X-tremeGENE or RNAiFect?? Is there a big difference compared to normal transfection reagents eg FuGene?? I am goin to transfect my epithelial cells in February and would like to test only the best reagents to save money and time.
Re: siRNA transfection
Posted by: MITian (IP Hidden, Unregistered user, )
Date: January 23, 2005 03:54PM
Hi Caro,
Yes, before you start your RNAi experiments, find a good transfection reagent and stick to it. I have used such reagent from ambion, invitrogen, Epoch Biolabs and have not compared side by side the effect. But the protocol for Invitrogen's lipofectamin 2000 and Epoch's GenCarrier seems much simpler (with GenCarrier is much cheaper) and that is why I later have only used it for all my transfections. Tom tuschl has some good comments on different transfection reagents: What about other transfection reagents? There are amazing differences in the efficiency of the cationic liposome reagents available on the market. Old, classical reagents are normally bad and only kept in the program of manufacturers to satisfy "conservative" customers, e.g. GenCarrier is about 10 times more efficient than the still distributed Lipofectene. We are just testing different transfection reagents.
Re: siRNA transfection
Posted by: RNAguy (IP Hidden, Unregistered user, )
Date: January 23, 2005 03:58PM
I've just tested two reagents from Ambion (siPORT Lipid and SiPORT Amine) side by side with Lipofectamine 2000 and GenCarrier. In my cells (a mouse beta-cell line) I got high levels of transfection using siPORT Amine, GenCarrier (>80% cells transfected), but the results with the other two lipidic reagents were dismal (approx 25% cells transfected). Other people in my lab gets 90% transfection levels with lipofectamine 2000 in fibroblasts cell lines. It all depends on the cell line. As MITian said, best thing to do is test several reagents at the beginning of your experiment and stick with the best.
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