Oneill Lab:Media: Difference between revisions
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#Adjust pH to 6.5 with KOH | #Adjust pH to 6.5 with KOH | ||
#Filter Sterilize | #Filter Sterilize | ||
==Links and Contacts== | |||
[[User:Seth|Seth Kasowitz]] [mailto:seth.kasowitz@uconn.edu] | |||
[http://www.oneill.mcb.uconn.edu/Michael/MONEILLhome.html|O'Neill Lab Homepage] |
Latest revision as of 13:09, 30 April 2007
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
- 10g Tryptone
- 5g Yeast Extract
- 10g NaCl
- Add deionized water to 1L
- Autoclave
Rich medium
- 10g Tryptone
- 5g Yeast Extract
- 5g NaCl
- 2g Glucose
- Add deionized water to 1L
- Autoclave
SOB
- 950ml Water
- 20g Tryptone
- 5g Yeast Extract
- 0.5g NaCl
- Dissolve
- Add 2.5ml 1M KCl
- Adjust pH to 7.0
- Add 10ml 1M MgCl2
- Autoclave
SOC
- Add 20ml sterile 1M Glucose to SOB
- Aliquot and freeze for storage
Media for Competent Cells
ψb
- 20g Tryptone
- 5g Yeast Extract
- 5g MgSO4
- Adjust pH to 7.6 with KOH
- Autoclave
TfbI
- Recipe for 500ml
- 1.47g Potassium Acetate
- 0.61g Rubidium Chloride
- 0.74g Calcium Chloride
- 4.95g Manganese Chloride
- Dissolve salts
- Add sterile glycerol to 15% (v/v)
- Adjust pH to 5.8 with 0.2M Acetic Acid
- Do not adjust pH with KOH
- Filter Sterilize
TfbII
- Recipe for 50ml
- 10mM MOPS or PIPES
- 0.15g PIPES
- 0.55g CaCl2
- 0.06g RbCl2
- 7.5ml Glycerol
- Adjust pH to 6.5 with KOH
- Filter Sterilize