Difference between revisions of "AU Biomaterials Design Lab:Protocols/Expression Culture Media"

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(New page: ==Description== Starter cultures enable us to get a jump-start on culturing bacteria to log-phase while we are expressing proteins. We will usually make 4 starter cultures for our expressi...)
 
 
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==Description==
 
==Description==
Starter cultures enable us to get a jump-start on culturing bacteria to log-phase while we are expressing proteins. We will usually make 4 starter cultures for our expressions.
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The expression culture will be able to support a large number of bacteria as they express our protein of interest. We will usually make 4 flasks-worth of expression culture.
  
 
==Protocol==
 
==Protocol==
# Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a 250mL Erlenmeyer flask
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*This needs to be done the day before protein expression
## 0.875g of LB
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# Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a 2.8L Fernbach flask
## 35mL of water
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## 25g of LB
 +
## 1L of water
 
# Cover the flask with foil
 
# Cover the flask with foil
# Autoclave
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# Autoclave (You can only fit two flasks into the autoclave at one time so you need to plan accordingly and perform two runs in the autoclave)
 
# Allow the flask to cool
 
# Allow the flask to cool
# Add 35uL of 1000X Antibiotic
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# Add 1mL of 1000X Antibiotic
# Inoculate with the bacteria you are culturing using one of the two following methods
 
## Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask
 
## Using sterile methods, add 5uL of a glycerol stock to the flask
 
# Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm
 
  
 
==Notes==
 
==Notes==
  
 
==References==
 
==References==

Latest revision as of 16:38, 11 August 2011

Description

The expression culture will be able to support a large number of bacteria as they express our protein of interest. We will usually make 4 flasks-worth of expression culture.

Protocol

  • This needs to be done the day before protein expression
  1. Prepare LB in a 2.8L Fernbach flask
    1. 25g of LB
    2. 1L of water
  2. Cover the flask with foil
  3. Autoclave (You can only fit two flasks into the autoclave at one time so you need to plan accordingly and perform two runs in the autoclave)
  4. Allow the flask to cool
  5. Add 1mL of 1000X Antibiotic

Notes

References