hi
i am traing to do stable lines with a plasmid i constructed, but having some troubles. the plasmid send a protein of intrest to the mitochondria.
i uses CT26 colon carcinoma and tried three diferent reagents- TFX, TransFast and fugene.
i am looking for a cell line that will transfect better, i dont mide if the sorse is human, mouse or rat, but that the mitochondrial activity is not known to be damaged.
thanks.
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4 replies to this topic
#2
laurequillo
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Posted 26 September 2010 - 07:30 AM
The efficiency in 293T is quite high. I dont know about the mitochondrial activity
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#3
virusfan
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Posted 26 September 2010 - 08:01 AM
shimrits, on 26 September 2010 - 01:14 AM, said:
hi
i am traing to do stable lines with a plasmid i constructed, but having some troubles. the plasmid send a protein of intrest to the mitochondria.
i uses CT26 colon carcinoma and tried three diferent reagents- TFX, TransFast and fugene.
i am looking for a cell line that will transfect better, i dont mide if the sorse is human, mouse or rat, but that the mitochondrial activity is not known to be damaged.
thanks.
i am traing to do stable lines with a plasmid i constructed, but having some troubles. the plasmid send a protein of intrest to the mitochondria.
i uses CT26 colon carcinoma and tried three diferent reagents- TFX, TransFast and fugene.
i am looking for a cell line that will transfect better, i dont mide if the sorse is human, mouse or rat, but that the mitochondrial activity is not known to be damaged.
thanks.
Hi there.
HeLa cells are quite easy to be transfected.
In between, which Fugene did you use? Fugene HD will transfect difficult-to-transfect cells better than Fugene6. Also, I would recommend jetPrime (http://www.polyplus-...ction-jetprime/). I am not advertising for the company. I found that it gives me better transfection efficiency of my endothelial cells than the transfection efficiency from other transfection reagents, such as Fugene HD, and Lipofectamine with PLUS reagent.
Good luck.
#5
rkay447
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Posted 28 September 2010 - 12:39 PM
I've made many stable cell lines with CHO-K1 cells (Chinese hamster ovary). They transfect very well and are easy to culture. Just a side note, perhaps your problems is not transfection efficiency though. Have you attempted to transfect a GFP vector into these cells and see no green cells? I tried to make a stable cell line that would express a domain and it turns out that the domain was highly toxic. I would see the cell line for about two passages before they all died.
Edited by rkay447, 28 September 2010 - 12:42 PM.
