Team:Calgary/Sandbox/Notebook/Protocols/OvernightCultures
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(Difference between revisions)
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<h2> Reagents and Materials (per culture prepared):</h2> | <h2> Reagents and Materials (per culture prepared):</h2> | ||
<ul> | <ul> | ||
- | <li> 10mL culture tube. Use 16mm x 160mm or 16mm x 125mm</li> | + | <li>10mL culture tube. Use 16mm x 160mm or 16mm x 125mm</li> |
- | <li> 5 mL LB</li> | + | <li>5 mL LB</li> |
- | <li> 5 μL 1000X antibiotics</li> | + | <li>5 μL 1000X antibiotics</li> |
- | <li> Single colonies on a plate (best not to start an overnight from a glycerol stock)</li> | + | <li>Single colonies on a plate (best not to start an overnight from a glycerol stock)</li> |
</ul> | </ul> | ||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
- | <li> Add 3mL sterile/autoclaved LB in a 15mL Falcon tube. </li> | + | <li>Add 3mL sterile/autoclaved LB in a 15mL Falcon tube. </li> |
- | <li> Pipet 3μL of 1000X antibiotic into the LB.</li> | + | <li>Pipet 3μL of 1000X antibiotic into the LB.</li> |
- | <li> Select a single colony using a sterile toothpick or pipette tip. </li> | + | <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> | + | <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> | + | <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 04:46, 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)