Quint Lab:yeast media

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muss bestellt werden 10x DO (Clontech) 1x: 0.6 g DO Supp + 26.7 g Min SD Base/ 1 l H2O 10x: 1.2 g DO Supp / 200 ml H2O → 121°C 20 min → 100 ml /1 l YSD component 400 ml 1 l 40% Dextrose/Glucose 160 g 400g → autoclave (or steril filtration) YPD liquid - no selection 400 ml 1 l Peptone/Tryptone 8 g 20 g Yeast Extract 4 g 10 g → pH 5.8 → add 380 ml 950 ml → autoclave 40 % Dextrose 20 ml 50 ml YPD plates - no selection 400 ml 1 l Peptone/Tryptone 8 g 20 g Yeast Extract 4 g 10 g Agar 8 g 20 g → pH 5.8 → add 380 ml 950 ml → autoclave 40 % Dextrose 20 ml 50 ml Y-SD liquid: (selection) Yeast Nitrogen Base w/o aa 2.7 g 6.7 g → pH 5.8 → add 340 ml 850 ml → autoclave 40 % Dextrose 20 ml 50 ml 10x Dropout 40 ml 100 ml (Trp, Leu, His oder Ade 4 ml 10 ml) Y-SD Plates: (selection) Yeast Nitrogen Base w/o aa 2.7 g 6.7 g Agar 8 g 20 g → pH 5.8 → add 340 ml 850 ml → autoclave 40 % Dextrose 20 ml 50 ml 10x Dropout 40 ml 100 ml (Trp, Leu, His oder Ade 4 ml 10 ml) 400 ml 1 l 10x Dropout: (selection) L-Isoleucine 120 mg 300 mg L-Valine 600 mg 1ֹ500 mg L-Arginine HCl 80 mg 200 mg L-Lysine 120 mg 300 mg L-Methionine 80 mg 200 mg L-Phenylalanine 200 mg 500 mg L-Threonine 800 mg 2ֹ000 mg L-Tyrosine 120 mg 300 mg L-Uracil 800 mg 200 mg → autoclave → store @ 4°C 100x Selection-aa L-Adenine hemisulphate salt 80 mg 200 mg L-Histidine HCl monohydrate 80 mg 200 mg L-Leucine 400 mg 1ֹ000 mg L-Tryptophan 80 mg 200 mg → autoclave → store @ 4°C 50% PEG: PEG 4000 200 g (Sigma #P-3640) → autoclave 10x TE: 1M Tris-HCl pH 7.5 100 ml 500mM EDTA pH 7.5 20 ml → add 880 ml → autoclave 10x Lithium Acetate: Lithium Acetate (Sigma #L-6883) 40.8 g → pH 7.5 with diluted acetic acid → autoclave PEG/LiAc dilution: (Transformation) 10 ml 150 ml 50% PEG 8 ml 120 ml 10x TE 1 ml 15 ml 10x LiAc 1 ml 15 ml → every time fresh!!!