Analysis Of Yeast Growth In YPD Agar

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Yeast Growth in YPD Agar:
To prepare YPD agar, mentioned in Table 1 nutrient ingredients in given concentration were weighed and added to 200 ml of distilled water. The mixture was autoclaved (SMS ASL80 MSV) for 1.5 hours at 121°C. On sterile plates, 25-30 ml of the mixture was poured and left to cool down. The yeast cells were then streaked on the agar plates and the cultures were grown in a stationary incubator (S1-600R, Lab Companion) for 72 hours at 30°C.
Yeast Growth in YPD:
To prepare YPD liquid medium, glucose, peptone and yeast extract were weighed and added to 200 ml of distilled water. The mixture was autoclaved for 1.5 hours at 121°C. Then a loop of yeast biomass from single cells grown on YPD agar was used to inoculate 5 ml of YPD. In order to disperse the yeast cells the content was vortexed briefly. The culture was grown in a shaker incubator (S1-600R, Lab Companion) for 24 hours at 30°C.
Lipid production:
For lipid production, 200 ml of mineral medium in glass bottle was inoculated with 10 ml of overnight culture in YPD to a final optical density at λ = 600 nm (OD600) of 0.2 and incubated on a shaker at 150 rpm and 30°C for 72 hours. Three mineral media were used (listed in materials section, Table 3).
6.2.2 Lipid droplet staining and microscopic observation
The stock solution was prepared by adding 1g of Oil Red O powder to 100 ml of isopropanol, in glass bottle. Prior to use, the working solution was prepared by diluting the stock solution with Mili Q water

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