yeast media
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
|
| YPD liquid - selection
| 400 ml
| 1 l
|
| Yeast Nitrogen Base w/o aa | 8 g | 20 g
|
| → pH 5.8
|
| → add | 380 ml | 950 ml
|
| → autoclave
|
| 40 % Dextrose | 20 ml | 50 ml
|
| 10x Dropout | 40 ml | 100 ml
|
| (Trp, Leu, His or Ade | 20 ml | 50 ml)
|
| YPD plates - selection
| 400 ml
| 1 l
|
| Yeast Nitrogen Base w/o aa | 8 g | 20 g
|
| Agar | 8 g | 20 g
|
| → pH 5.8
|
| → add | 380 ml | 950 ml
|
| → autoclave
|
| 40 % Dextrose | 20 ml | 50 ml
|
| 10x Dropout | 40 ml | 100 ml
|
| (Trp, Leu, His or Ade | 20 ml | 50 ml)
|
| 10x dropout
| 400 ml
| 1 l
|
| L-Isoleucine | 120 mg | 300 mg
|
| L-Valine | 600 mg | 1500 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 | 2000 mg
|
| L-Tyrosine | 120 mg | 300 mg
|
| L-Uracil | 800 mg | 2000 mg
|
| → autoclave
|
| → store @ 4°C
|
| 100x Selection-aa
| 400 ml
| 1 l
|
| L-Adenine hemisulphate salt | 80 mg | 200 mg
|
| L-Histidine HCl monohydrate | 80 mg | 200 mg
|
| L-Leucine | 400 mg | 1000 mg
|
| L-Tryptophan | 80 mg | 200 mg
|
| → autoclave
|
| → store @ 4°C
|
50% PEG (for 400 ml):
(Sigma #P-3640)
→ autoclave
10x TE (for 1 l):
- 1M Tris-HCl pH 7.5 - 100 ml
- 500mM EDTA pH 7.5 - 20 ml
→ add 880 ml ddH2O
→ autoclave
10x Lithium Acetate (for 400 ml):
- Lithium Acetate (Sigma #L-6883) - 40.8 g
→ pH 7.5 with diluted acetic acid
→ autoclave
| PEG/LiAc dilution for 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!!!
|
