Isolation of bioluminescent bacteria
Goal
In the ocean many organisms produce light. There are also many diffent colors. In this practical we will isolate these bacteria from fresh fish.
Materials
- 1 Fresh fish
- 4 Artificial Seawater plates, simply add 10% agar before autoclaving
- 100 mL Artificial Seawater
- 2 50 mL Erlenmeyer flasks
- Glass spatula
- Inoculation loop
- 100 mL beaker glass with some 95% ethanol
- Gas burner
- Air pump, tube and diffuser
Method
- Sterilize a inoculation loop in a flame
- When it is cooled down, scrape the loop along the skin of the fish
- Streak the loop onto the plate
- Repeat this procedure onto a second plate
- Incubate one plate at RT(24° C) and one in the fridge (8° C)
- Take a look at your plates in total darkness after 2 days or more
- As soon as you can observe colonies, restreak these on fresh plates
- As soon as all colonies look the same, you may assume you have a pure culture
- Sub culture a colony in a 25 mL ASW liquid medium in a Erlenmeyer flask. Incubate at 24° C.
- Put the tube with diffuser in the flask and turn on the air pump
Back to Biofactory - Class 1: Incubator