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Team:Cambridge/Protocols/Plates
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Revision as of 10:16, 14 August 2012
Contents |
Making LB Agar plates
Preparation
Prepare liquid broth (LB) from liquid broth capsules and distilled water.
Making LB Agar (15%)
- 1. In a 1 litre bottle, add 3.75g of Agar powder to 250ml LB
- 2. Autoclave to sterilize
- 3. These can be stored and heated up to make plates when needed
Antibiotics
Different antibiotics will have different optimum concentrations
- Chloramphenicol (25mg/ml)
Add 250mg to Chloramphnicol to 10ml ethanol (100%). Use 1ml for 1 litre of LB Agar to make final concentration 25ug/ml plates.
Making LB Agar Plates
- 1. Cool down/heat up LB agar to approx. 55 degrees Celsius
- 2. In a flow hood, add 250ul antibiotics to 250ml LB agar; mix
- 3. Pour approx. 25ml into each plate
- 4. Wait approx. 30 mins for plates to set
- 5. Store in 4 degrees Celsius fridge for future use
