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

Plate Codes

Line color Media Antibiotic Black only LB none Black, Red LB Amp Black, Blue LB Tet Black, Green LB Kan Black, Cyan 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

Working concentration 20 ug/ml (4 ml stock solution per liter)

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 – 25 mg/ml

Weigh out 1.25 g Chloramphenicol (Sigma C0378) Add 50 ml 100% Ethanol Mix/vortex vigorously

Working concentration 25 ug/ml (1 ml stock solution per liter)