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.

1 Fresh fish, make sure it is really fresh from the sea. Ask for it at a fish shop.
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

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