media and transfection - change? (Mar/31/2008 )
This might sound like a simple question but I am confused. I have been doing transfections for awhile now and use the lipid based transfection method. I used to change the media 24 hours post transfection and use to even plate my cells in antibiotic free media since the stress to the cells would be too great during transfection. With some newer transfection reagents they clearly say that you dont need to use antibiotic free media when plating your cells and I also have not needed to change the media post transfection. I was wondering if this method is not advisable and/or why????? Many thanks in advance...
can you tell by your own observation? look for cell morphology, target gene expression with positive treatment and control groups?
Morphology looks the same im just worried about gene expression. I just started not changing media so I dont know about that. I just dont want to go through all the troubleshooting, lol. Thanks though
Morphology looks the same im just worried about gene expression. I just started not changing media so I dont know about that. I just dont want to go through all the troubleshooting, lol. Thanks though
this depends on which cell line will be used. For example, if you use HEK293, you need not change the medium. the best way is doing transfection in normal medium. if you find cell death, then try to use p/s free medium when doing transfection. the next day change the medium with normal medium. besides, which transfection reagent you use is also important. commonly, lipofectamin could give you high transfection efficiency but more toxic to cells. So if you use lipofectamin, p/s free medium is recommanded. if you use fugene 6, you need not do that.
Hello,
Some transfection reagent request to use complete medium , and you do not need change the medium for all your experiement time.No antibiotics add during your experiement .If you ,see some change in your cells ,you should optimize your experiement by change transfection reagent or reduce it concentration and DNA concentration too.I would like to advice you , to use OptiMEM or any medium reduced serum for tranfect your gene ,then change the medium 12 to 24 hrs post-tranfection into complete medium.I want to know ,what kind of cells do you use and transfection reagent .Note that some transfection reagent if you used it in requested amounts it might be toxic to the cells and even kill the cells.
Ela