Transfection of Hi5 cells with Fugene

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Tue Apr 26 16:26:25 EST 2011


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


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