MinD+ is used for growing yeast to use with the luminometer. This media has been shown to give 5-10x higher GCN4-FLuc/RLuc ratios than YEPD. MAKE SURE YOUR BOTTLE LABELS VERY CLEARLY REFLECT WHETHER OR NOT GLUCOSE AND/OR AMINOACIDS HAVE BEEN ADDED, AND THE DATE THEY WERE ADDED TO THE MIN.
How to make MinD+NOTE: It is best to divide up one liter into 4, 500mL bottles with 250mL of solution. Add the glucose and amino acids only to the bottle that you are currently using and parafilm the ones you are not using. This gives you back up bottles of media in case of contamination.
SD medium (Hinnebuch's minimal) - makes 1L
6.7g yeast nitrogen base (without amino acids but with ammonium sulfate) Product number: Difco 291940
To each bottle with 250mL of SD medium:
2.5mL 50mM Valine (bio-world.com Catalog # 42290000-2)
2.5mL 50mM Isoleucine (bio-world.com Catalog # 40900012-3)
2.5mL 200mM Lysine (bio-world.com Catalog # 41200044-2)
2.5mL 200mM Histidine (bio-world.com Catalog # 4080052)
5ml 100mM Leucine (Sigma Catalog # L8000-250G)
How to make amino acid solutions:
Valine- aka "100X"
0.2929g in 50mL sterile water
Isoleucine- aka "100X"
0.3280g in 50mL sterile water
Lysine- aka "100X"
1.4619g in 50mL sterile water
Histidine- aka "100X"
1.556g in 50mL sterile water
Leucine- aka "50X"
.656g in 50mL sterile water
Steriflip (filter sterilize 0.22um) all amino acid solutions and store at 4 degrees C
Clearly label on the bottle with tape when each the glucose and amino acids are added.