UA Biophysics:Protocols:SD 1L

Stocks

Yeast Nitrogen Base(YNB) 10X stock 1L

• 67 g YNB
• Fill the bottle with MilliQ until 1L
• Autoclave

Dropout(DO) Supplement 10X stock 1L

• 7.5 g CSM -His-Ura
• Fill the bottle with MilliQ until 1L
• Autoclave

Carbon Source(CS) 10X stock 1L (May be Dextrose or Galactose)

• Autoclave two 1L Schott
• 200 g CS
• Dilute CS in Schott with MilliQ until 1L. May take several minutes at room temperature and shaking
• 'Filter with 0.22 um filter into the other Schott

Essential Amminoacid(EA) Supplements 10X stocks 1L. (if the yeast strain has His or Ura auxotrophy)

• Autoclave two 1L Schott
• 200 mg Histidine
• 200 mg Uracil
• Dilute in Schott with MilliQ until 1L
• Filter with 0.22 um filter into the other Schott

Mix 100 mL of all the components. Complete to 1L steril distilled water

No Stocks

Another way to make SD medium is without preparing concentrated stocks. This is useful for turbidostat use. In a 500 mL erlenmeyer flask

• 6700 mg YNB
• 750 mg DO Supplement
• Fill with MilliQ until 500 mL
• Autoclave

In another 500 mL erlenmeyer flask

• Fill with milliQ 500 mL
• Autoclave

Wait until it cools, then add

• 20 g Carbohidrate (D-Glucose or D-Galactose)
• 20 mg Uracil
• 20 mg Histidine
• dilute and filter with 0.22 um filter into the YNB flask

Please, Do not hesitate to contact me if you have any question Juan David Estupiñán

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