2x YEP + Glu/Gal
Ingredient |
amount |
final conc. |
|---|---|---|
Yeast extract |
20g |
20g/L |
Peptone |
40g |
40g/L |
20% Glu/Gal |
200 mL |
4% |
ddH20 |
800 mL |
|
It's better to make 2x YEP - sugar media and 20% sugar media separately and add them to make the complete media just before setting up the cultures.
Sunrise Media for Yeast
Note: "YNB+Nitrogen": This can be confusing, because different companies define "YNB" differently. At Sunrise Science Products, they define "YNB" as the salts, vitamins and trace elements that form a base to which nitrogen can be added. Unless theproduct name says "+Nitrogen", there is no nitrogen in the product. Read other company labels carefully to be sure you are replacing the item correctly with theirs.
Sunrise Yeast Nitrogen Base (YNB) contains salts, vitamins and trace elements as defined: Potassium phosphate (1000 mg/L), Inositol (2 mg/L), Copper sulfate (0.04 mg/L), Magnesium sulfate (500 mg/L), Niacin (0.4 mg/L), Potassium iodide (0.1 mg/L), Sodium chloride (100 mg/L), PABA (0.2 mg/L), Ferric chloride (0.2 mg/L), Calcium chloride (100 mg/L), Pyridoxine (0.4 mg/L), Manganese sulfate (0.4 mg/L), Biotin (0.002 mg/L), Riboflavin (0.2 mg/L), Sodium molybdate (0.2 mg/L), Pantothenate, calcium (0.4 mg/L), Thiamine (0.4 mg/L), Zinc sulfate (0.4 mg/L), Folic acid (0.002 mg/L), Boric acid (0.5 mg/L).
Synthetic Dropout Media Recipe (if you are using Sunrise's YNB + Nitrogen bottle)
Name |
Final Concentration |
For 500 mL Volume |
|---|---|---|
Yeast Nitrogen Base + Ammonium Sulfate (YNB + Nitrogen) |
6.7 g/L |
3.35 g |
Amino acid (AA) mix (CSM-AA) |
As noted on the container |
x g |
ddH2O |
|
450 mL |
Yeast Agar (for plates only) |
20 g/L |
10 g |
AUTOCLAVE AND THEN COOL, THEN ADD... |
|
|
Sterile dextrose (20% w/v stock concentration) |
2% w/v |
50 mL |
Synthetic Dropout Media Recipe (if you are using Sunrise's YNB bottle):
Name |
Final Concentration |
For 500 mL Volume |
|---|---|---|
Yeast Nitrogen Base (YNB) |
1.7 g/L |
0.85 g |
Ammonium Sulfate |
5 g/L |
2.5 g |
Amino acid (AA) mix (CSM-AA) |
As noted on the container |
x g |
ddH2O |
|
450 mL |
Agar (for plates only) |
20 g/L |
10 g |
AUTOCLAVE AND THEN COOL, THEN ADD... |
|
|
Sterile dextrose (20% w/v stock concentration) |
2% w/v |
50 mL |
CSM mix for making drop out aa mix:
CSM Component |
mg/L (1x) |
g/L (100X) |
Adenine |
10 |
1 |
L-Alanine |
0 |
0 |
L-Arginine |
50 |
5 |
L-Asparagine |
0 |
0 |
L-Aspartic Acid |
80 |
8 |
L-Cysteine |
0 |
0 |
Glutamine |
0 |
0 |
L-Glutamic Acid |
0 |
0 |
Glycine |
0 |
0 |
L-Histidine |
20 |
2 |
Myo-Inositol |
0 |
0 |
L-Isoleucine |
50 |
5 |
L-Leucine |
100 |
10 |
L-Lysine |
50 |
5 |
L-Methionine |
20 |
2 |
Para-AminoBenzoic Acid (PABA) |
0 |
0 |
L-Phenylalenine |
50 |
5 |
L-Proline |
0 |
0 |
L-Serine |
0 |
0 |
L-Threonine |
100 |
10 |
L-Tryptophan |
50 |
5 |
L-Tyrosine |
50 |
5 |
Uracil |
20 |
2 |
L-Valine |
140 |
14 |
Remove AAs from this mix to make corresponding drop out mixes (e.g. to make -Ura-Trp drop out, add all the above AAs except Ura and Trp)
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