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Team:Calgary/Sandbox/Notebook/Protocols/OvernightCultures
From 2013.igem.org
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| Line 13: | Line 13: | ||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
| - | <li>Add 3mL sterile/autoclaved LB in a 15mL Falcon tube | + | <li>Add 3mL sterile/autoclaved LB in a 15mL Falcon tube</li> |
| - | <li>Pipet 3μL of 1000X antibiotic into the LB | + | <li>Pipet 3μL of 1000X antibiotic into the LB</li> |
| - | <li>Select a single colony using a sterile toothpick or pipette tip | + | <li>Select a single colony using a sterile toothpick or pipette tip</li> |
| - | <li>Place toothpick or pipette tip in the culture tube and stir | + | <li>Place toothpick or pipette tip in the culture tube and stir</li> |
| - | <li>Remove toothpick, or in the case of a pipette tip, leave in the tube | + | <li>Remove toothpick, or in the case of a pipette tip, leave in the tube</li> |
<li>Place culture tube in incubator at 37 degrees Celsius overnight shaking vigorously (250 RPM)</li> | <li>Place culture tube in incubator at 37 degrees Celsius overnight shaking vigorously (250 RPM)</li> | ||
</ol> | </ol> | ||
Revision as of 04:54, 16 September 2013
Overnight Cultures
Reagents and Materials (per culture prepared):
- 10mL culture tube. Use 16mm x 160mm or 16mm x 125mm
- 5 mL LB
- 5 μL 1000X antibiotics
- Single colonies on a plate (best not to start an overnight from a glycerol stock)
Protocol
- Add 3mL sterile/autoclaved LB in a 15mL Falcon tube
- Pipet 3μL of 1000X antibiotic into the LB
- Select a single colony using a sterile toothpick or pipette tip
- Place toothpick or pipette tip in the culture tube and stir
- Remove toothpick, or in the case of a pipette tip, leave in the tube
- Place culture tube in incubator at 37 degrees Celsius overnight shaking vigorously (250 RPM)
