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
Min D
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

1L sterile water

Autoclave for 30-45 min then ADD 10mL OF STERILE 50% GLUCOSE per 250mL of solution.

To each bottle with 250mL of SD medium:

2.5mL 50mM Valine ( Catalog # 42290000-2)

2.5mL 50mM Isoleucine ( Catalog # 40900012-3)

2.5mL 200mM Lysine ( Catalog # 41200044-2)

2.5mL 200mM Histidine ( 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.