21.07.

2014 1

Preparation of Yeast Media

YP 10 g/l yeast extract (1 % w/v)

20 g/l peptone (2 % w/v)
20 g/l glucose or galactose (2 % w/v)
80 mg/l adenine sulfate

A yeast extract 10 g
Peptone 20 g
Adenine sulfate stock solution (table page 2) 20 ml
Ad 900 ml H2Odd
Autoclave

B Glucose or galactose stock solution (20 % w/v) 100 g ad 500 ml H2Odd

Autoclave

For use: combine A + B (1/10 of final volume), i.e. 900 ml A + 100 ml B

 Final volume: 1000 ml YP-Medium

Glucose or galactose is autoclaved separately to avoid caramelisation.

YP agar plates ( 25 ml/ small plate)

A yeast extract 5g
Peptone 10 g
Adenine sulfate stock solution (table page 2) 10 ml
Agar (20 g/l = 2 % w/v) 10 g
Ad 450 ml H2Odd
Autoclave, cool to 70 °C, do not forget the stirrer bar!

B Glucose or galactose stock solution (20 % w/v) 100 g ad 500 ml H2Odd

Autoclave

For use: combine A + B (1/10 of final volume), i.e. 450 ml A + 50 ml B (add slowly)
 Final volume: 500 ml.
Pour plates.
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Minimal Medium (SC) Nitrogen base, as specified by supplier

5 g/l ammonium sulfate
20 g/l glucose or galactose (2 % w/v)

A yeast nitrogen base 1.9 g

Ammonium sulfate 5g
Ad 900 ml H2Odd
Autoclave

B Glucose or galactose stock solution (20 % w/v) 100 g ad 500 ml H2Odd

Autoclave

For use: combine A + B (1/10 of final volume), i.e. 900 ml A + 100 ml B

 Final volume: 1000 ml SC-Medium. Add the appropriate markers (WHALU).

SC agar plates ( 25 ml/ small plate)

A yeast nitrogen base 0.95 g

Ammonium sulfate, (NH4)2SO4 2.5 g
Ad 250 ml H2Odd
Autoclave, cool to 70 °C, do not forget the stirrer bar!

B Agar (20 g/l = 2 % w/v) 10 g

Ad 200 ml H2Odd
Autoclave

C Glucose or galactose stock solution (20 % w/v) 100 g ad 500 ml H2Odd

Autoclave

For use: combine A + B + C (1/10 of final volume), i.e. 250 ml A + 200 ml B + 50 ml C (add
slowly) and Marker (WHALU) as required  Final volume: 500 ml.
Pour plates.

For 150 ml Stock solution

Marker
Marker-Mix (for single use)
Adenine sulfate A, Ade 600 mg 400 mg / 100 ml
Uracil U, Ura 150 mg 200 mg / 100 ml
L-Tryptophane W, Trp 300 mg 1 g / 100 ml
L-Histidine H, His 200 mg 1 g / 100 ml
L-Leucine L, Leu 450 mg 1 g / 100 ml
For Marker-Mix: weigh, mix and autoclave any combination of aminoacids you need (use
1 ml per 50 ml medium). Like the other components the stock solutions of the markers or
marker mixes are autoclaved and can be stored. L-Tryptophane is stored in the dark
(aluminium foil), and need to be warmed to solve.