Wittrup: E. coli Media: Difference between revisions
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'''Ampicillin''' | '''Ampicillin''' | ||
Stock: 25 mg/mL of sodium salt of ampicillin (amp) in water. | *Stock: 25 mg/mL of sodium salt of ampicillin (amp) in water. | ||
*Filter sterilize and store in aliquots of 4 mL at -20 oC | |||
Filter sterilize and store in aliquots of 4 mL at -20 oC | *Working concentration = 35-50 mg/L for liquid media. Often 100 mg/L for plates. | ||
Working concentration = 35-50 mg/L for liquid media. Often 100 mg/L for plates. | |||
'''SOC Media''' | '''SOC Media''' | ||
Revision as of 12:40, 10 February 2006
LB Media
Liquid, 1000 mL
- 10g Bacto tryptone
- 5g Bacto yeast extract
- 10g NaCl
pH to 7.5 with NaOH (optional?) Add ddH2O to final volume. Filter sterilize, or autoclave. Add antibiotic if selective media is desired (perhaps 100 mg/L amp), but do not add until cooled to below 50 oC (touchable by bare hand)
Plates, 1000 mL
- Ingredients as above (still save antibiotic for last)
- 15-18g agar
- Autoclave. Add antibiotic if selective media is desired, but do not add until cooled to below 50 oC (touchable by bare hand)
Ampicillin
- Stock: 25 mg/mL of sodium salt of ampicillin (amp) in water.
- Filter sterilize and store in aliquots of 4 mL at -20 oC
- Working concentration = 35-50 mg/L for liquid media. Often 100 mg/L for plates.
SOC Media
100 mL
- 2.0g 2% Bactotryptone
- 0.5g 0.5% Yeast extract
- 10 mM NaCl (0.2 mL 5 M)
- 10 mM MgCl2 (1.0 mL 1 M)
- 10 mM MgSO4 (1.0 mL 1 M)
- 20 mM Dextrose (D-glucose) (0.36 g)
Add ddH20 to final volume. Filter sterilize, or autoclave.
