Lipofection of 293 cell line with large plasmid DNA; help req.

[Jack Guida]

Obaid,

We have been using lipofectin from GIBCO/BRL for some time now with good 
results, make sure that you dont have any antibiotics in the media during 
the transfection.  Also, don't leave the lipofectin on for more than 12 
hours, the 293s are very sensitive to it.  You could check this paper for 
more information:  Biotechniques 15:868-872 (1993).  Good Luck

  Jack D. Guida             
  Dept. of Microbiology and Immunology         
  Albert Einstein College of Medicine
  guida at aecom.yu.edu        

On Tue, 5 Dec 1995, O.Khan wrote:

> I am attempting to transfect 293 cells with large ~40kb plasmid carrying the 
> adenovirus whole genome, using DOTAP. I dont seem to get good efficiency of 
> transfection. Also I get a high cell death, (DOTAP-DNA-media mixture is kept for 
> overnight on the cells). Can anyne having experience with lipofecting large plasmids 
> help me out with this.
> Thanks,
> Obaid Yusuf Khan
> 
> 
>