| YPD
(complete medium) 10 g Yeast extract
20 g Peptone 20 g Agar 900 ml dH2O Autoclave separately 20 g glucose 100 ml dH2O for plates add one NaOH pellet SD (dropout medium, complemented synthetic medium) 6,7 g yeast nitrogen base without
amino acids
0,6 g aa mix without His, Trp, Leu, Ura (CSM) 20 g agar nn mg amino acids/uracil (powder) 900 ml dH2O autoclave separately 20 g sugar (glucose or galactose or galactose+10 g raffinose for plates add one NaOH pellet Indicator plates (to assay ß-gal activity) 3,35 g yeast nitrogen base
without amino acids
0,3 g aa mix without His, Trp, Leu, Ura (CSM) 10 g agar nn mg amino acids/uracil (powder) one pellet of NaOH 400 ml dH2O 50 ml 1 M K-phosphate, pH 7 10 g sugar 50 ml H2O autoclave separately at 55°C add 1 ml X-Gal (20 mg dissolved in 1 ml DMF), mix before pouring the plates store plates at 4°C not longer than two months amino acids/uracil: 20 mg/l uracil (100x stock 2,4 mg/ml) 20 mg/l histidine (100x stock 2,4 mg/ml) 60 mg/l leucine (100x stock 7,2 mg/ml) 40 mg/l tryptophane (100x stock 5 mg/ml) fill powder into a 2l flask (two fold volume), add NaOH pellet (prevents degradation of agar) and stir bar, add water (1x deionised), autoklave, prepare separately sugar solution or other supplements. Cool down flasks in a water bath to 55°C, mix and pour plates. Dry plates for one to two days on the bench. plates and media can be stored at room temperature, although for longterm storage 4°C are recommended. |