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Protocol/2
From 2010.igem.org
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* 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. | ||
| - | * Add 5mL non-contaminated LB. Then add antibiotic | + | * Add 5mL non-contaminated LB. Then add antibiotic. |
* Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture. | * Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture. | ||
* Place culture tube in incubator at 37<sup>o</sup>C overnight shaking vigorously (250 rpm). | * Place culture tube in incubator at 37<sup>o</sup>C overnight shaking vigorously (250 rpm). | ||
Revision as of 01:03, 26 October 2010
Protocol 2: 5 mL Overnight
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.
