User:Klare Lazor/Notebook/Chem-496-001/2013/03/04

Objective

• Create agar plates 20
• Started bacteria culture
• Autoclaved all solutions

Description

Autoclave

• 8 Flask of 50mL of water and agar.

1. Prepare LB in a 250mL Erlenmeyer flask
2. 0.875g of LB
3. 35mL of water
4. Cover the flask with foil
5. Autoclave
6. Allow the flask to cool
7. Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm

• Separate for starter culture
• Make certain that there is enough water in the reservoir before you begin.

1. Turn the power on
2. Turn the black knob to fill. This fills the chamber with water.
3. Let water continue to fill the chamber until it reaches the line at the front of the chamber.
4. Turn the black knob to STE (short for "sterilize").
5. Load your media into the autoclave chamber. (Note: 2 2.8L Fernbach Flasks can be loaded into the chamber)
6. Close the autoclave door and seal it shut.
7. Set the temperature knob just to the right of 121F.
8. Turn the timer knob to 50 minutes.
9. After the 50 minutes have elapsed, wait 30 minutes until the pressure drops back to atmospheric pressure.
10. Turn the black knob to Off.
11. Use gloves to unload your sterilized material.

Starter Culture

1. Inoculate with the bacteria you are culturing using one of the two following methods
2. Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask
3. Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm

Data

• Add data and results here...

Notes

This area is for any observations or conclusions that you would like to note.

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