(a) Minimal Medium (pH 7.1):

Prepare a Fe2(SO4)3 solution by adding 40 mg of Fe2(SO4)3 to 50 ml of double-distilled water (ddH2O)

Add 1 L ddH2O to a 2 L glass beaker

Add the following components to the beaker:

13.61 g KH2PO4

1.98 g (NH4)2SO4

4.2 g KOH

0.2 g MgSO4

1 ml of the Fe2(SO4)3 solution

50 mg L-leucine

50 mg L-histidine

50 mg adenine

20 mg L-arginine-HCl

20 mg L-methionine

30 mg L-tyrosine

30 mg L-isoleucine

30 mg L-lysine-HCl

25 mg L-phenylalanine

100 mg L-glutamic acid

150 mg L-valine

375 mg L-serine

Put the beaker on heated stirrer with a magnetic flea and stir until it is all dissolved

Check that the pH is 7.1 and adjust if necessary

Using a 50 ml sterile syringe dispense 45 ml aliquots into glass bottles with metal screw top lids

Sterilize the Minimal Medium at 121 °C for 10 min in an autoclave

Store at room temperature

(b) D-(+)-Glucose:

Prepare a 20% w/v solution in ddH2O

Dispense 20 ml aliquots in to glass vials with metal screw top lids

Sterilize the Glucose solution at 121 °C for 10 min in an autoclave

Store at room temperature

(c) L-Aspartic Acid:

Make a stock solution of 4 mg/ml in ddH2O

Dispense 20 ml aliquots in to glass vials with metal screw top lids

Sterilize the L-Aspartic Acid solution at 121 °C for 10 min in an autoclave

Store at room temperature

(d) Vitamin Solution:

Prepare a biotin solution by adding 2 mg of biotin to 100 ml of ddH2O

Weigh out 8 mg thiamine, 8 mg pyridoxine, 8 mg pantothenic acid, 40 mg inositol. Add all the dry components and 20 ml of the biotin solution to 180 ml ddH2O

Make 10 ml sterile aliquots by filtering through a 0.2-µm pore size disposable filter into sterile glass bottles, in a laminar air flow cabinet

Store at 4 °C

(e) L-Threonine:

Prepare 100 ml of 24 mg/ml L-threonine in ddH2O

Dispense 10 ml aliquots in to glass vials with metal screw top lids

Sterilize the L-threonine solution at 121 °C for 10 min in an autoclave

Store at room temperature

(f) Copper(II) Sulfate:

Prepare 25 ml of a 20 mM Copper(II) Sulfate solution in ddH2O

Make 5 ml sterile aliquots by filtering through a 0.2-µm pore size filter into sterile glass bottles, in a laminar flow cabinet

Store at room temperature

(g) Chlorophenol red-β-D-galactopyranoside (CPRG):

Prepare 25 ml of a 10 mg/ml solution of CPRG in ddH2O

Make 5 ml sterile aliquots by filtering through a 0.2-µm pore size filter into sterile glass bottles, in a laminar flow cabinet

Store at 4 °C