Dear Pepa, do you have access to a quicker positive control like GFP? If you don't have a fluorescence microscope, beta galactosidase or - glucuronidase and similar are suitable genes, too, as long as you can get the appropriate X-substrate. The big advantage against westerns is that you also will get an idea of the transfection efficiency, as you will see single transfected cells.
Do you have the possibility to test your transfection reagent with some other cell line to check if it is ok? Are your cells healthy?any mycoplasma? really Hi5 cells? (some paranoia might be ok :) You also may dig the archives of this NG for polyethyleneimine (PEI, or simply find it here: http://www.bio.net/bionet/mm/methods/1999-December/080050.html). You only need to get the right one, it's crucial (see the protocol) and it's worth a try for sure I think, as it's horribly cheap: You'll get a whole bathtub full of final transfection reagent which is sufficient for a whole flock of sheep (see http://forums.biotechniques.com/viewtopic.php?f=18&t=13968 for details, it also works on cows), for a price much less than 1 single commercial kit). BTW, it's at least similar to JetPEI. Other hopefully useful links are in this post here: http://www.bio.net/bionet/mm/methods/2002-May/093201.html Have fun! Wo _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
