Growth media for Saccharomyces cerevisiae

YPD medium (Ausubel et al., 1992)

• 1% Select Yeast extract (Gibco BRL) 10 g
• 2% Bacto-peptone (Difco Laboratories) 20 g
• 2% glucose. 100 ml 20 % Glucose

Yeast extract and peptone (YP) have to be prepared in 9/10 volumes of water and autoclaved. 20 % Glucose solution is autoclaved separately. At room temperature, 1/10 vol glucose solution is added to the YP solution.

• 1% Select Yeast extract (Gibco BRL) 10 g
• 2% Bacto-peptone (Difco Laboratories) 20 g
• 4 mg/l adenine 2 ml of a 2 mg/ml stock solution
• 20 mg/l tryptophan 4 ml of a 5 mg/ml stock solution
• 2% glucose. 100 ml 20 % Glucose

Yeast extract, peptone, adenine and tryptophan (YP⁺) have to be prepared in 9/10 volumes of water and autoclaved. 20 % Glucose solution is autoclaved separately. At room temperature, 1/10 vol glucose solution is added to the YP solution.

YPR medium

• 2% Bacto-peptone
• 1% Select Yeast extract
• 4% raffinose

YPGal

• 2% Bacto-peptone
• 1% Select Yeast extract
• 2% galactose

YPGE

• 2% Bacto-peptone
• 1% Select Yeast extract
• 3% glycerol
• 2% ethanol

• 0.67% Select Yeast nitrogen base without amino acids (Difco Laboratories) 6,7 g
• 0.08% complete supplement mixture minus uracyl (Bio101, Vista, CA) 0,8 g
• 2% glucose 100 ml 20% glucose

Solutions of yeast nitrogen base and complete supplement on the one hand, and the glucose stock solution on the other hand, have to be autoclaved separately.

Wach et al. suggest to add 50 mM NaH₂PO₄ pH 6,4 (= 15,6 g NaH₂PO₄ + 40 ml 1M NaOH + 160 ml H₂O) if the culture will be used to prepare DNA.

• 0.67% Select Yeast nitrogen base without amino acids (Difco Laboratories) 6,7 g
• 0.08% complete supplement mixture minus uracyl (Bio101, Vista, CA) 0,8 g
• 2% glucose 20 g

Glucose, together with 15 g BiTek Difco agar (in case of solid medium), are dissolved in 900 ml of deionised water and autoclaved. YNB (w/o AA and (NH₄)₂SO₄), (NH₄)₂SO₄ and the required amino acids/bases are solubilized in 100 ml and autoclaved separatly.

! +6: to be added for SD+6 plates
! +13: to be added (together with +6) for SD+13 plates
! For SD+16, all ingredients have to be added; for adenine, 10 ml instead of 2 ml stock solution should be added.

! Tryptophane (Sterile filtrated) and histidine have to be stored at 4°C in the dark

! ^‡ Sterile filtration; should be added to the medium after having autoclaved the medium.

! For selective medium, do not add the components on which there will be selected for.

• 0.17% Select Yeast nitrogen base without amino acids and ammoniumsulphate (Difco Laboratories) 1,7 g
• 0,5 % Ammoniumsulphate 5 g
• 50 mg/l Uracil 25 ml stock solution (2 mg/ml)
• 75 mg/l Adenine 37,5 ml stock solution (2 mg/ml)
• 75 mg/l Leucine 7,5 ml stock solution (10 mg/ml)
• 20 mg/l Tryptophane 4 ml stock solution (5 mg/ml)
• 30 mg/l Lysine 3 ml stock solution (10 mg/ml)
• 20 mg/l Histidine 2 ml stock solution (10 mg/ml)

Combine in a solution of 150 ml and autoclave

• 2% glucose 20 g
• 2% Difco Bacto Agar 20 g

Solubilize in 850 ml water and autoclave, together with a magnetic stirrer.

Combine the two solutions, and add 1 g of 5-FOA
Mix the solution gently at approx. 60°C up to complete solubilization of the 5-FOA.

Solid media are prepared by adding 2% Select Agar (Gibco BRL) to liquid medium. 20 g

Autoclaved medium is supplemented, as required, with

• 200 mg/l G-418 (Geneticin^R sulfate from Gibco/BRL; Germany)
  ! the activity of this chemical may be batch-dependent !)