Chemicals: campesterol (~98%) and ergosterol (≥75%) are obtained from Sigma-Aldrich. β-Sitosterol (~65%) and stigmasterol (~95%) are purchased from Fisher Scientific. All engineered yeast strains in this work are listed in Table S1 and constructed in a haploid CENPK2.1D background (MATα; his3D;1 leu2-3_112; ura3-52; trp1-289; MAL2-8c; SUC2). Yeast strains were cultured at 30 o C in complex yeast extract peptone dextrose (YPD, all components from BD Diagnostics) medium or synthetic defined medium (SDM) containing yeast nitrogen base (YNB) (BD Diagnostics), ammonium sulfate (Fisher Scientific), 2% (w/v) glucose unless specified and the appropriate dropout (Takara Bio) solution for selection. 200 mg/L G418 sulfate (Calbiochem) or 200 mg/L Hygromycin B (Life Technologies) were used in YPD medium for selection. Plasmids used in this study are in Table S5. Sequences of genes used in this work are listed in Table S6.
Complex yeast extract peptone dextrose ypd
Manufactured by BD
Complex yeast extract peptone dextrose (YPD) is a nutrient-rich media formulation commonly used for the growth and cultivation of a variety of microorganisms, particularly yeasts. It provides the necessary carbon, nitrogen, and other essential nutrients required for microbial proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Ammonium sulfate, by Thermo Fisher Scientific (2 mentions)
Yeast nitrogen base, by BD (2 mentions)
Campesterol, by Merck Group (1 mentions)
Hygromycin b, by Thermo Fisher Scientific (1 mentions)
β sitosterol, by Thermo Fisher Scientific (1 mentions)
Ergosterol, by Merck Group (1 mentions)
Lysogeny broth medium, by Thermo Fisher Scientific (1 mentions)
2 protocols using complex yeast extract peptone dextrose ypd
1
Engineered Yeast Strains for Sterols
2
Yeast and Bacterial Culture Conditions
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A haploid CENPK2.1D (MATα; his3D;1 leu2-3_112; ura3-52; trp1-289; MAL2-8c; SUC2) was used in this work. Yeast strains were cultured at 30 o C in complex yeast extract peptone dextrose (YPD, all components from BD Diagnostics) medium or synthetic defined medium (SDM) containing yeast nitrogen base (YNB) (BD Diagnostics), ammonium sulfate (Fisher Scientific), 2% (w/v) unless specified and the appropriate dropout (Takara Bio) solution for selection. E. coli strain TOP10 was used in the inhibition assay, and cultured at 37 °C in lysogeny broth (LB) medium (Fisher Scientific).
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