Bacterial Media Recipes

Contents

1. Super Optimal Broth (SOB)
2. Super Optimal broth with Catabolite repression (SOC)
3. Terrific Broth (TB)
4. 2XYT
5. YES Media
6. SC Media

1. Super Optimal Broth (SOB)

Component	Amount	Final concentration
NaCl	0.5 g
tryptone	20 g
deionized H2O	to 950 ml
yeast extract	5 g

1. Shake until the solutes have dissolved.
2. Add 10 ml of a 250 mM solution of KCl. (This solution is made by dissolving 1.86 g of KCl in 100 ml of deionized H2O.)
3. The pH of the medium should be near pH 7.0. Check and adjust with 5 N NaOH.
4. Adjust the volume of the solution to 1 liter with deionized H2O.
5. Sterilize by autoclaving for 20 minutes on liquid cycle.
6. Just before use, add 5 ml of a sterile solution of 2 M MgCl2. (This solution is made by dissolving 19 g of MgCl2 in 90 ml of deionized H2O. Adjust the volume of the solution to 100 ml with deionized H2O and sterilize by autoclaving for 20 minutes at 15 psi [1.05 kg/cm2] on liquid cycle.)

Recipe from CSH Protocols

2. Super Optimal broth with Catabolite repression (SOC)

• Protocol

Note: the pH needs to be adjusted to 7.0 with NaOH (~200 uL of 5 N NaOH), which is not mentioned in the above protocol.

3. Terrific Broth (TB)

Used mainly for growing high densities of DH5a or similar cells to produce high concentrations of plasmid DNA. Recipe taken from CSH protocols

Make the following separately:

3.1 Salts Solution

Final concentration: 0.17 M KH2PO4, 0.72 M K2HPO4

1. This solution is made by dissolving 23.1 g of KH2PO4 and 125.4 g of K2HPO4 in 900 mL of H2O.
2. After the salts have dissolved, adjust the volume of the solution to 1000 mL with H2O and filter the solution.

3.2 TB solution

Component	Amount	Final concentration
Deionized H2O	to 900 mL
Tryptone	12 g
Yeast extract	24 g
Glycerol	4 mL

1. Shake until the solutes have dissolved.
2. Sterilize by autoclaving for 20 min on liquid cycle.
3. Allow the solution to cool to 60°C or less, and then add 100 mL of a sterile salts solution from above.

4. 2XYT

Component	Amount	Final concentration
Bacto Tryptone	16g
Bacto Yeast Extract	10g
NaCl	5g

1. Adjust pH to 7.0 with 10N NaOH.
2. Adjust to 1L using distilled H20.
3. Sterilize by autoclaving.

5. YES Media

Component	Amount	Final concentration
ddH2O	to 1L
YES media	36.13 g

1. Stir for 10 minutes.
2. Use a 0.2 uM membrane filter (disposable filter) and attach it to the bottle near an open flame.
3. Vacuum filter sterilization into a 1L bottle.
4. Detach the filter next to the flame and flame the cap before placing it onto the bottle.

6. SC Media

Component	Amount	Final concentration
ddH2O	to 1L
drop out mix synthetic minus uracil without yeast nitrogen	1.92 g
yeast nitrogen base without amino acid	6.7 g
glucose	20 g	2%

1. Heat with stirring until dissolved.
2. Adjust to desired pH if needed.
3. Sterilize with 0.2 uM membrane filter (disposable filter).

Note: unless making SC-leu, be sure to add 225 mg/L leucine to these when using -leu dropout mix if the SC powder does not contain it already