Oneill Lab:Media

Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology

Recipes for making media commonly used in lab

Bacterial growth media

• Recipes are for 1L liquid media
• Can use recipes to make plates by adding bacto agar to concentration of 1.5% w/v
• Top agar is used by adding bacto agar to 0.7% w/v

LB

1. 10g Tryptone
2. 5g Yeast Extract
3. 10g NaCl
4. Add deionized water to 1L
5. Autoclave

Rich medium

1. 10g Tryptone
2. 5g Yeast Extract
3. 5g NaCl
4. 2g Glucose
5. Add deionized water to 1L
6. Autoclave

SOB

1. 950ml Water
2. 20g Tryptone
3. 5g Yeast Extract
4. 0.5g NaCl
5. Dissolve
6. Add 2.5ml 1M KCl
7. Adjust pH to 7.0
8. Add 10ml 1M MgCl₂
9. Autoclave

SOC

1. Add 20ml sterile 1M Glucose to SOB
2. Aliquot and freeze for storage

Media for Competent Cells

ψb

1. 20g Tryptone
2. 5g Yeast Extract
3. 5g MgSO₄
4. Adjust pH to 7.6 with KOH
5. Autoclave

TfbI

• Recipe for 500ml

1. 1.47g Potassium Acetate
2. 0.61g Rubidium Chloride
3. 0.74g Calcium Chloride
4. 4.95g Manganese Chloride
5. Dissolve salts
6. Add sterile glycerol to 15% (v/v)
7. Adjust pH to 5.8 with 0.2M Acetic Acid
   • Do not adjust pH with KOH
8. Filter Sterilize

TfbII

• Recipe for 50ml

1. 10mM MOPS or PIPES
   • 0.15g PIPES
2. 0.55g CaCl₂
3. 0.06g RbCl₂
4. 7.5ml Glycerol
5. Adjust pH to 6.5 with KOH
6. Filter Sterilize

Links and Contacts

Seth Kasowitz [1]

Lab Homepage