20/08/13

From 2013.igem.org

Making nutrient agar plates and broth

• Making up broth and agar solution to grow P. putida in.
• For agar plates:
  • 9.2g of nutrient agar medium
  • 400ml of distilled water
• For broth:
  • 3.2g of nutrient broth
  • 400ml of distilled water
• Both were autoclaved for 45mins
• Agar was poured onto 16 plates

Plating out bacteria

• Freeze dried P. putida was rehydrated using 1ml of nutrient broth
• The culture was aseptically transferred back into the broth containing tube
• Mix
• From that tube, colonies were streaked onto 6 plates and left to incubate over night at 30C

Growing bacteria in broth

• 100ml of autoclaved broth was added to 2 flasks each
• The broth tube, containing the bacteria, was divided between flasks
• Flask were left to incubate at 30C on a shaker overnight