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Team:Cambridge/Protocols/Plates
From 2012.igem.org
(Difference between revisions)
| Line 18: | Line 18: | ||
===Making LB Agar (15%)=== | ===Making LB Agar (15%)=== | ||
| - | 1. In a 1 litre bottle, add 3.75g of Agar powder to 250ml LB | + | :1. In a 1 litre bottle, add 3.75g of Agar powder to 250ml LB |
:2. Autoclave to sterilize | :2. Autoclave to sterilize | ||
:3. These can be stored and heated up to make plates when needed | :3. These can be stored and heated up to make plates when needed | ||
| Line 29: | Line 29: | ||
===Making LB Agar Plates=== | ===Making LB Agar Plates=== | ||
| - | 1. Cool down/heat up LB agar to approx. 55 degrees Celsius | + | :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 | :2. In a flow hood, add 250ul antibiotics to 250ml LB agar; mix | ||
:3. Pour approx. 25ml into each plate | :3. Pour approx. 25ml into each plate | ||
:4. Wait approx. 30 mins for plates to set | :4. Wait approx. 30 mins for plates to set | ||
:5. Store in 4 degrees Celsius fridge for future use | :5. Store in 4 degrees Celsius fridge for future use | ||
Revision as of 15:59, 13 July 2012
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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. 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
