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