AU Biomaterials Design Lab:Protocols/Starter Culture Media: Difference between revisions
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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. | 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. | ||
==Protocol== | ==Protocol== | ||
# Prepare LB in a 250mL Erlenmeyer flask | # Prepare [[AU_Biomaterials_Design_Lab:Materials/LB|LB]] in a 250mL Erlenmeyer flask | ||
## 0.875g of LB | ## 0.875g of LB | ||
## 35mL of water | ## 35mL of water | ||
Latest revision as of 15:54, 11 August 2011
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.
Protocol
- Prepare LB in a 250mL Erlenmeyer flask
- 0.875g of LB
- 35mL of water
- Cover the flask with foil
- Autoclave
- Allow the flask to cool
- Add 35uL 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
