description: 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).
attributes.tag: isolate
attributes.value: ATCC CRL-1573
attributes.tag: age
attributes.value: fetus
attributes.tag: biomaterial_provider
attributes.value: ATCC CRL-1573
attributes.tag: sex
attributes.value: female
attributes.tag: tissue
attributes.value: embryonic kidney
attributes.tag: BioSampleModel
attributes.value: Human