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Protocol/2
From 2010.igem.org
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Protocol 2: 5 mL Overnight | Protocol 2: 5 mL Overnight | ||
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* 10 mL culture tube Use 16mm x 160mm or 16mm x 125 mm | * 10 mL culture tube Use 16mm x 160mm or 16mm x 125 mm | ||
Revision as of 00:53, 26 October 2010
Protocol 2: 5 mL Overnight
Reagents:
- 10 mL culture tube Use 16mm x 160mm or 16mm x 125 mm
- 5 mL 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 5 mL non-contaminated LB. Do this first, 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 37 C overnight shaking vigorously (250 rpm).
Notes:
For I0500, culture overnight; then add 1 mM ITPG in morning and continue culture for another 3 hrs. Then miniprep.
