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Protocols
Media
LB
Ingredients:
10 g Bacto-tryptone
5 g yeast extract
10 g NaCl
SOB
Ingredients
0.5% (w/v) yeast extract
2% (w/v) tryptone
10 mM NaCl
2.5 mM KCl
20 mM MgSO4
Per liter
5 g yeast extract
20 g tryptone
0.584 g NaCl
0.186 g KCl
2.4 g MgSO4
SOC
Ingredients
SOB
20 mM glucose
CCMB80 buffer for preparation of chemically competent cells
Materials
Detergent-free, sterile glassware and plasticware
Table-top OD600nm spectrophotometer
SOB
CCMB80 buffer
10 mM KOAc pH 7.0 (10 ml of a 1M stock/L)
80 mM CaCl2.2H2O (11.8 g/L)
20 mM MnCl2.4H2O (4.0 g/L)
10 mM MgCl2.6H2O (2.0 g/L)
10% glycerol (100 ml/L)
Adjust pH DOWN to 6.4 with 0.1N HCl if necessary
Adjusting pH up will precipitate manganese dioxide from Mn containing solutions
Filter sterilize and store at 4°C
Slight dark precipitate appears not to affect its function
