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Good quality frozen stock can be used.
Either way, frozen stock will keep for weeks or months, though its flavor will deteriorate somewhat over time.
From the frozen stock, blood agar plates were inoculated and incubated at 37 °C to obtain cultures.
Pre-cultures were done by inoculating 100 µl from frozen stock tubes into 5 ml of TSB and then incubated at 20, 30 or 37 °C.
A primary potato dextrose broth culture was inoculated with Conidiospores from a −80 °C frozen stock and grown at 28 °C, shaking at 200 rpm for 2 days.
One of the additional simulations used frozen stock of isolate cultures, the other used frozen stock of isolate cultures with glycerol added to a final concentration of 10%.
Cultures were plated and grown overnight from frozen stock.
KHT cell cultures were established from viably frozen stock tumour pieces.
These frozen stock cultures were used to inoculate precultures for all fed-batch experiments.
All experiments were performed using cells in exponential growth phase from passage 5 to 20 post-thawing from frozen stock.
All cultures used in this study had undergone a maximum of 2 in vitro passages after recovery from frozen stock.
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