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Seeds were cultured on standard media, Phytamax Orchid Maintenance Media (Sigma-Aldrich Chemical, St Louis, MO, USA) with 0.7%% (w/v) agar.
Media were replaced with maintenance media with antibiotics 24 h later after transfection, and then, nickel was added to the media.
Mouse ESC was differentiated by removing leukemia inhibitory factor (LIF) from the maintenance media in the presence or absence of the specific DNA methyltransferase (DNMT) inhibitor 5′-aza-2′-deoxycytidine (aza-dC).
Cardiomyocytes were seeded with CDI's plating media into fibronectin (5 µg/mL) coated wells at the appropriate density (2.4e5 cells per well of 12 well plate; 2e4 per well of 96 well plate. After 2 days, media was changed to CDI's maintenance media and replaced every other day.
From day 4 to day 8, cells were kept in maintenance media with new maintenance media being replaced by every two days.
The DNA complex was then removed and replaced with neuronal maintenance media.
Induction media for the adipogenic differentiation contained dexametasone, IBMX and insulin, and maintenance media was with insulin only.
On day 4, progression media was replaced with maintenance media (DMEM supplemented with 10% fetal calf serum and penicillin/streptomycin).
Adipogenic differentiation was assayed after incubation in induction media for 6 days and 3 days in maintenance media.
The neuronal maintenance media was removed from the cells and they were washed with phosphate buffered saline (PBS) containing no calcium or magnesium.
For cell viability/toxicity assays, cells were grown in a Phenol Red-free media that was otherwise identical in composition to the regular maintenance media.
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