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huh-7 transfection efficiency


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#1 SF_HK

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Posted 14 August 2013 - 11:57 PM

I'm trying to transfect huh-7 cell line but I'm getting very low GFP tranefction efficiency after 24 hours of transfection. Only 1-2% of cells are transfected with GFP. I've used liopfectamine LTX reagent + Plus transfection reagent.

 

My cells were 60% confluent at the time of transfection as this cell lines grows very fast I wanted to avoid very dense cell po[ulation prior to transfection. WHat I did observe that single cells were not transfected but cells growing as a colony of 2 or more cells had a greated uptake of GFP?  Has anyone experienced soemthing similar?  If this is the case then perhaps I should wait for the cells to be about 80% confluent on the day of tranfection and grown in colonies instead of single cells.  I've heard huh-7 cells are hard to transfect. ANy advice would eb appreciated.






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