Wittrup: E. coli Media: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
'''LB Media''' | '''LB Media''' [''see also [[LB]]''] | ||
''Liquid, 1000 mL'' | ''Liquid, 1000 mL'' | ||
| Line 12: | Line 12: | ||
*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''' | '''Ampicillin''' [''see also [[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''' [''see also [[SOC]]''] | ||
''100 mL'' | ''100 mL'' | ||
Latest revision as of 14:38, 13 February 2006
LB Media [see also LB]
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 [see also 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 [see also SOC]
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.
