Download Selective Yeast Culture Induction

Scharenberg Lab Protocol: Selective Yeast Culture & Induction
Submitted by: Jordan Jarjour
Date Updated: 1/04/2008
SC media (1L) (Synthetic complete drop out media + selected amino acids)
 Yeast nitrogen base without amino acids (Sigma #Y0626) 6.7 g/L
 Yeast synthetic drop-out media supplement without Tryptophan (Sigma #Y1876)
1.92g/L; or without Uracil (Sigma #Y1501) 1.92g/L; or without Trp, Ura, His, Leu
(Sigma #Y2001) 1.40g/L, then add back amino acids as necessary…ie. His and Leu can
be added at 76mg/L and 380mg/L to make –Trp –Ura media or plates.
 pH to 5.6 with NaOH
 900mL total volume (ddH2O)
o For plates, add 20g/L bacteriological agar
 Autoclave (20’ sterilization step).
o For plates, add 1/10 volume of 20% Glucose (AFTER AUTOCLAVING)
o Add Penicillin (100 i.u./ml), Streptomycin (100g/ml) and Kanamycin (25g/ml)
Listed below are concentrations for complete drop-out medium (mg/L):
Adenine hemisulfate
18
Alanine
76
Arginine HCl
76
Asparagine monohydrate
76
Aspartic acid
76
Cysteine HCl monohydrate
76
Glutamic acid monosodium salt
76
Glutamine
76
Glycine
76
Myo-inositol
76
Histidine
76
Isoleucine
76
Leucine
380
Lysine monohydrochloride
76
Methionine
76
p-Aminobenzoic acid potassium salt
8
Phenylalanine
76
Proline
76
Serine
76
Threonine
76
Tryptophan
76
Tyrosine disodium salt
76
Uracil
76
Valine
76
Immediately before use add carbon source:
 2% Raffinose (w/v) and 0.1% Glucose for growth without induction

2% Galactose to induce expression (maximal expression at 6-8hrs post induction)