D:\533564743.doc 15 March 2007
LB Medium- solid
Ingredients:
LB Broth- 25 grams/ Liter
1.5% Bactoagar ( Difcoagar)-15grams/Liter
preparation:
Add all the above ingredients in 1 liter of dd water and mix them thoroughly (with stir
bar). check the pH with the pH indicator paper. The pH should be 7.5.
Autoclave for 121 degree Celsius for 30 minutes in liquid cycle.
Cool down to 60 degree Celsius. Then, add antibiotics.
LB Medium- Liquid
Ingredients:
LB Broth - 25grams/liter
preparation:
Add all the above ingredients in 1 liter of dd water and mix them thoroughly (with stir
bar). check the pH with the pH indicator paper. The pH should be 7.5.
Autoclave for 121 degree Celsius for 30 minutes in liquid cycle.
Cool down to 60 degree Celsius. Then, add antibiotics. If you want to store in 4 degree
for long time . please don’t add antibiotics. You could add antibiotics ahead of time.
Make it fresh always. Lb with antibiotics is not good if you store more than one month.
Antibiotics Concentration
Final concentration to LB medium.
Kanamycin- 50ug/ml
Amplicilin-50 to 100ug/ml
HygromycinTetracycline- 10ug/ml
SpectinomycinStocks
Kanamycin- 50mg/ml, so 1ul has 50ug of kanamycin
Amplicilin-100mg/ml
HygromycinTetracycline-10mg/ml in 70% ETOH
Spectinomycin-150mg/ml of water
D:\533564743.doc 15 March 2007
Glycerol storage solution (2x) 100 ml
LB
70 ml
30% glycerol
30 ml
autoclave at 121C for 20-30 min
autoclave tubes, too.
Add 1 vol of this solution to your E. coli liquid culture (e.g. add 500 l storage solution
to 500 l cell culture). Mix well. Store at –80 C.