Bacterial Media
Missouri Western Synthetic Biology Protocols Bacterial Media Prep
LB Broth
5 g Yeast Extract
10 g Tryptone
5 g NaCl
QSV 1 liter
LB Plates (makes about 35 plates)
5 g Yeast Extract
10 g Tryptone
5 g NaCl
15 g BactoAgar
QSV 1 liter
SOC Broth
2 g Bacto Tryptone
0.5 g Bacto Yeast Extract
0.2 mL 5M NaCl
0.25 mL 1M KCl
1 mL 1M MgCl2
1 mL 1M MgSO4
2 mL 1M Dextrose (Glucose)
90 mL of dH2O
Mix, then fill up to 100 mL with dH2O
Plate Codes
- Black line only LB none
- Black line, Red line LB Amp
- Black line, Blue line LB Tet
- Black line, Green line LB Kan
- Black line, Cyan line LB Chlor
Ampicillin
Stock – 50 mg/ml
Weigh out 2.5 g Ampicillin, sodium salt (Sigma A0166) Dissolve in 25 ml dH20 Add 25 ml Ethanol
Working concentration 50 ug/ml (1 ml stock solution per liter)
Tetracycline
Stock – 5 mg/ml
Weigh out 0.25 g Tetracycline HCl (Sigma T7660)
Dissolve in 50 ml 70% Ethanol
Mix/vortex vigorously
Filter sterilize
Make 4 ml aliquot in foil covered 15 ml tubes
Working concentration 20 ug/ml (4 ml stock solution per liter)
Light Sensitive! Keep plates in darkness
Kanamycin
Stock – 10 mg/ml
Weigh out 0.5 g Kanamycin sulfate (Sigma K1377) Dissolve in 50 ml dH20
Working concentration 20 ug/ml (2 ml stock solution per liter)
Chloramphenicol
Stock – 35 mg/ml
Weigh out 1.75 g Chloramphenicol (Sigma C0378) Add 50 ml 100% Ethanol Mix/vortex vigorously
Working concentration 35 ug/ml (1 ml stock solution per liter)
L-Arabinose
Stock – 250 mg/ml
Weigh out 2.5 g L-Arabinose Add 10 ml dH2O Mix/vortex vigorously
Working concentration 0.5 mg/ml (2 ml stock solution per liter)
