HEK 293 with transfection of an empty pCI plasmid (HEK-pCI)
HEK 293 cells were maintained by the procedures of Bulanenkova et al., 2007. T25 flasks with approximately 10^6 HEK 293 cells were transfected using Lipofectamine 3000 (Invitrogen, cat#L3000015, Carlsbad, CA, USA) according to manufacturer's recommendations. Briefly, immediately before transfection, media was changed on the flask. Lipofectamine:DNA complexes for the pCI null vector (Promega, cat#E1731, Madison, WI, USA) were added to the media with 6.5ug DNA administered. Lipofectamine:DNA complexes were prepared in OptiMEM medium (Gibco, cat#31985062, Gaithersburg, MD, USA), then added to HEK growth media (RPMI 1640:DMEM:F-12 (2:1:1) medium supplemented with 10% fetal bovine serum [Gibco: RPMI-1640: cat#11875093; DMEM/F12: cat#11320033; FBS: cat#A3160402]) currently in the flask. Following transfection, flasks were returned to incubate at 37 C /5% CO2. At 48 hrs post infection, the transfection medium was replaced with fresh HEK growth medium in each flask. At 72 hrs post infection, flasks were harvested by scraping cells in PBS, and DNA was isolated by a Wizard Genomic DNA Purification Kit (Promega, cat#A1120).
isolate
ATCC CRL-1573
age
fetus
biomaterial_provider
ATCC CRL-1573
sex
female
tissue
embryonic kidney
BioSampleModel
Human