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suspension cell transient transfection


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#1 labrat612

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Posted 05 March 2010 - 01:19 PM

Hey guys!

I'm trying desperately to get a handle on this, but every time I try, I fail. So, basically I'm transiently transfecting suspension CHO cells that are in spinner flasks. I'm currently using Fugene HD as the transfection reagent-- and so far no luck.
To compound this, the plasmid I'm using does not have a marker on it. So I have to wait to run either an ELISA or a western blot for my protein that should be secreted into the media.

Any ideas on a protocol that might work?? I'm losing my mind (or what is left of it).

~Labrat 612

#2 KevinK

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Posted 05 March 2010 - 01:59 PM

Labrat612,
Could you post some details on your transfection protocol? The FUGENE HD reagent should be working quite well on CHO cells in suspention. You can review the protocol we have at Promega in our database here: http://www.promega.c...tools/FugeneHD/

With some details on your procedure I may be able to make some suggestions.

Thanks,
Kevin
Promega Corporation
Madison, WI

#3 mostafa

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Posted 06 March 2010 - 02:14 AM

HI before you lose your mind it is better you use some plasmid having reporter gene like EGFP with your construct. this is so cheeper and easier to do.




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