Method prepared by Grace Panganiban: gepangan@facstaff.wisc.edu


Yeast Dropout Powder

Use 1.6 g of thoroughly mixed powder/liter
Use a spatula to make sure different grain sizes are represented
Can make without specific amino acids for selection purposes, but need to recalculate g/l.

Adenine 1.25 g
Arginine 0.9
Aspartate 3.0
Glutamate 3.0
Lysine 0.9
Methionine 0.6
Phenylalanine 1.5
Serine 11.25
Threonine 6.0
Tyrosine 0.9
Valine 4.5
Alanine 1.2
Asparagine 1.2
Cysteine 1.2
Glutamine 1.2
Glycine 1.2
Isoleucine 1.2
Proline 1.2
Histidine 0.6 - 100x=200 mg/100 ml
Leucine 1.8 - 100x=600 mg/100 ml
Tryptophan 1.2 - 100x=400 mg/100 ml
Uracil 0.6 - 100x=200 mg/100 ml

Rich medium/YDP

10 yeast extract
20 g peptone
20 g dextrose
diw to 1 liter

No need to pH; yeast like acidic conditions

Complete minimal medium

1.3 g of appropriate dropout medium
6.7 g of YNB-AA/AS
5 g NH4(SO4)4
(20 g agar)
diw to 900 mls

Add 100 mls of sterile 20% glucose (or galactose) after autoclaving
Yeast grow almost as well in this medium as they do in YDP