Wittrup: E. coli Media: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 5: | Line 5: | ||
*5g Bacto yeast extract | *5g Bacto yeast extract | ||
*10g NaCl | *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, but do not add until cooled to below 50 oC (touchable by bare hand) | 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'' | ''Plates, 1000 mL'' | ||
| Line 11: | Line 11: | ||
*15-18g agar | *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) | *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''' | '''SOC Media''' | ||
Revision as of 12:39, 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.
