LB Broth - 1L

1. To 950 mL of deionized water, add:
   • Tryptone 10 g
   • Yeast Extract 5 g
   • NaCl 10 g
2. Fill to 1L. Autoclave for 1 hour.

SOB - 1L

1. To 950 mL of deionized water, add:
   • Tryptone 20 g
   • Yeast Extarct 5 g
   • NaCl 0.5 g
2. Shake until the solutes have dissolved. Add 10 mL of a 250 mM solution of KCl (dissolve 0.465 g KCl in 25 mL water). adjust pH to 7.0 using NaOH. Adjust the volume of the solution to 1 liter with deionized water. Sterilize by autoclaving for 20 minutes.
3. Just before use, add 5 mL of a sterile solution of 2 M MgCl2, which can be made by dissolving 19 g of anhydrous MgCl2 in 90 mL of deionized water (adjust mass accordingly if using MgCl2 hexahydrate) and autoclaving for 20 minutes.

SOC

SOC medium is identical to SOB medium, except it also contains 20 mL glucose. After the SOB medium has been autoclaved, allow it to cool to 60 degrees or less and then add 20 mL of a sterile 1 M solution of glucose (dissolve 90 mL of dI water, adjust volume to 100 mL, then filter sterilize).

M9 Supplemented Media - 1L

• 200 mL 5X minimal salts - make 1 L of salts by dissolving the following in dI water to final volume of 1 L, divide into 200 mL aliquots, and autoclave for 15 minutes.

Na2HPO4*7H2O	64 g
KH2PO4	15 g
NaCl	2.5 g
NH4Cl	5 g

• 34 mL of 10mg/mL thiamine - dissolve 10 mg per mL of water and filter-sterilize
• 10 mL of 40% glycerol - add 80 mL glycerol to 120 mL of water and autoclave to sterilize (can also use 40% glucose)
• 20 mL of 10% Casamino acids - dissolve 50 g of Bacto Casamino acids in 500 mL water and autoclave to sterilize
• 2 mL of 1M MgSO4 - dissolve 24.65 g MgSO4*7H2O in 100 mL water and autoclave to sterilize
• 100 uL of 1M CaCl2 - dissolve 14.7 g of CaCl2*2H2O in 100 mL of water and autoclave to sterilize
• Autoclave and sterilize 734 mL of dI water.
• Combine above solutions using sterile technique, adding sterile water at the end to bring final volume up to 1L. There may be some precipitation during preparation but precipitate should go back into solution once the volume is brought up to 1L. Add antibiotic as appropriate and store at 4 degrees C.