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Polybrene was added at a concentration of 8 µg/ml and the HepG2 cells were incubated with this medium over night to induce HNF6 expression.
For spore germination experiments, mating colonies were grown in glutamate medium over night at 25°C, washed with 30% ethanol, washed with water, and then incubated in minimal medium at 32°C in the absence of uracil.
BMMCs were supplied with fresh culture medium over night to ensure maximum viability.
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Let it dry over night.
The plasmid born CA3427 gene was over-expressed in E. coli BL21 in 1L flasks containing TB medium over one night at 17°C after induction with IPTG (500 µM) at OD600 nm = 0.5.
Then the medium must equilibrate over night on a thermo plate at 37°C under atmospheric air.
Single yeast colonies were inoculated in selective SD medium and incubated over night at 30°C.
BL21 transformant colonies were inoculated in 100 ml of LB/ampicillin medium and incubated over night at 37°C.
After that, the Transwell insert was inverted in a 24 well cell culture cluster (Costar, Corning Incorporated) in DMEM complete medium and placed over night at 37°C in humidified atmosphere with 5% CO2.
The cells were allowed to attach for 4 5 hours, thereafter the medium and the non-attached cells were aspirated and fresh medium added for culture over night.
The aorta was cut in 1 2 mm thick rings, and all the rings were cultured in serum-free Optimem medium (Invitrogen, Basel, Switzerland) over night.
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