Team:Calgary/Sandbox/Notebook/Protocols/OvernightCultures

From 2013.igem.org

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<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>5mL LB</li>
-
<li>5 μL 1000X antibiotics</li>
+
<li>5μL 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>

Latest revision as of 01:25, 18 September 2013

Overnight Cultures

Reagents and Materials (per culture prepared):

  • 10mL culture tube. Use 16mm x 160mm or 16mm x 125mm
  • 5mL LB
  • 5μL 1000X antibiotics
  • Single colonies on a plate (best not to start an overnight from a glycerol stock)

Protocol

  1. Add 3mL sterile/autoclaved LB in a 15mL Falcon tube
  2. Pipet 3μL of 1000X antibiotic into the LB
  3. Select a single colony using a sterile toothpick or pipette tip
  4. Place toothpick or pipette tip in the culture tube and stir
  5. Remove toothpick, or in the case of a pipette tip, leave in the tube
  6. Place culture tube in incubator at 37 degrees Celsius overnight shaking vigorously (250 RPM)

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