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Team:Alberta/Notebook/protocols/overnight
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| - | Protocol | + | ==Protocol: 5mL Overnights== |
| - | Reagents: | + | ===Reagents:=== |
* 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm | * 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm | ||
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| - | Procedure: | + | ===Procedure:=== |
* Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube. | * Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube. | ||
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| - | Notes: | + | ===Notes:=== |
For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep. | For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep. | ||
Revision as of 00:13, 27 October 2010
Protocol: 5mL Overnights
Reagents:
- 10mL culture tube Use 16mm x 160mm or 16mm x 125 mm
- 5mL LB
- Concentrated antibiotics
- Single colonies on a plate (Best not to start O/N from glycerol stocks)
Procedure:
- Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
- Add 5mL non-contaminated LB. Then add antibiotic.
- Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture.
- Place culture tube in incubator at 37oC overnight shaking vigorously (250 rpm).
Notes:
For I0500, culture overnight; then add 1mM ITPG in morning and continue culture for another 3 hours. Then miniprep.
