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The inoculated PDA media were transferred in quantity of 20 ml in sterilized Petri dishes (d = 9 cm) and allowed to solidify.
Aliquots of the culture media were transferred to 96-well microplates.
1.0×106 cells in 100 µl of media were transferred in duplicate wells to a 96-well plate.
Equal volumes of the media were transferred to non-treated sister cultures, which were further incubated for either 1 or 24 hours.
Inoculum plugs (3 mm diameter removed from the growing margins of colonies from identical MMN media) were transferred to the cellophane-covered agar in the Kilner jars.
Aliquots of culture media were transferred to 96-well microplates.
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20 ml of nutrient agar media was transferred into petriplates and they were left undisturbed for 1-2 h.
The overnight culture then centrifuge and re-suspended in appropriate media, was transferred into Erlenmeyer flask containing 1 L cultures of each media and were incubated at 37 °C, at 225 rpm.
To perform the transfection 50 µl of serum free media was transferred into a micro-centrifuge tube; 3 µl of the FuGene 6 reagent was then added directly to this and mixed by gentle flicking.
50 μL of media was transferred to a white plate and combined with the luciferin agent.
Thereafter, the media was transferred to osteoinductive media with 10−8 M dexamethasone for a further 7 days.
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