Media
Daniel Melamed 15/08/07
YPD
For 1 liter:
Yeast extract – 10 gr
Bacto peptone – 20 gr
Glucose – 20 gr
Supplement with water and autoclave.
For plates – Dissolve all the above ingredients in 500 ml water and autoclave. Mix
with autoclaved 500 ml water containing 15 gr Agar and pour.
YNB
For 1 liter:
Yeast nitrogen base with out amino acids and ammonium sulfate – 1.7 gr
Ammonium sulfate – 5 gr
Threonine – 0.35 gr
Dissolve all ingredients in 900 ml water and autoclave.
Add pre-autoclaved 100 ml of 20% glucose and necessary amino acids to make SD
(synthetic dropout) media.
For plates – dissolve the above ingredients in 400 ml water and autoclave. Mix with
pre-autoclaved 100 ml of 20% glucose and 500 ml of water containing 15 gr Agar.
Add necessary amino acids and pour.
Amino acids and nucleotides for 1 liter medium:
Methionine 20 mg/ml – 2 ml
Adenine 5 mg/ml – 8 ml
Lysine 10 mg/ml – 5 ml
Uracil 2 mg/ml – 10 ml
Tryptophan 10 mg/ml – 5 ml
Histidine 20 mg/ml – 1 ml
Leucine 10 mg/ml – 8 ml
Tyrosine 2 mg/ml – 15 ml
We routinely prepare 100 ml of each amino acid stock solution, dissolve and
dispense it to three or four 50 ml tubes and autoclave. The amounts can be calibrated
also for striking required amino acids on pre-made YNB plates (25 ml). We typically
prepare MALY plates (YNB containing methionine, adenine and lysine) and add the
appropriate amino acids regarding the yeast strain and the plasmids that are used.
LB
For 1 liter:
Tryptone – 10 gr
NaCl – 10 gr
Yeast extract – 5 gr
For plates only- add 15 gr Agar
Autoclave
When possible we prefer to divide the liquid media to 100-200 ml aliquots since this
medium tend to contaminate.