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

1. Black line only LB none
2. Black line, Red line LB Amp
3. Black line, Blue line LB Tet
4. Black line, Green line LB Kan
5. 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)