Difference between revisions of "Bacterial Media"
| Line 40: | Line 40: | ||
Weigh out 0.25 g Tetracycline HCl (Sigma T7660) | Weigh out 0.25 g Tetracycline HCl (Sigma T7660) | ||
| + | |||
Dissolve in 50 ml 70% Ethanol | Dissolve in 50 ml 70% Ethanol | ||
| + | |||
Mix/vortex vigorously | Mix/vortex vigorously | ||
| + | |||
Filter sterilize | Filter sterilize | ||
| + | |||
Make 4 ml aliquot in foil covered 15 ml tubes | Make 4 ml aliquot in foil covered 15 ml tubes | ||
Revision as of 15:43, 5 October 2010
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
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 C7795) Add 50 ml 100% Ethanol Mix/vortex vigorously
Working concentration 35 ug/ml (1 ml stock solution per liter)
