Dose anyone use OptiMem for cell transient transfection.
Do you know how long I can keep BHK cells in this Medium untill changing it to DMEM:F12 supp with FCS
(BHK cells were transiently transfected with wild-type and mutant β-glucuronidase cDNAs in pcDNA3.1(+) or with vector alone by using 8 μl Lipofectamine and 2 μg plasmid DNA in a total volume of 200 μl OptiMEM in 35-mm dishes. After a 5-h incubation, medium was replaced with DMEM containing 5% FCS and penicillin/streptomycin.)
hi gsovak,
according to
Storch et al., 2003 (Mutational analysis in longest known survivor of mucopolysaccharidosis type VII)
you can leave them at least for 5 hours in optimem
I guess that should be long enough for a transfection
http://www.springerlink.com/content/avr1qjkxa16mmuxh/fulltext.pdf
(BHK cells were transiently transfected with wild-type and mutant β-glucuronidase cDNAs in pcDNA3.1(+) or with vector alone by using 8 μl Lipofectamine and 2 μg plasmid DNA in a total volume of 200 μl OptiMEM in 35-mm dishes. After a 5-h incubation, medium was replaced with DMEM containing 5% FCS and penicillin/streptomycin.)
Thanks Vanishing,
I am doing a 12 incubation with Opitimem after transfection and it is working now.
Guy