Team:British Columbia/Notebook/Protocols/media

From 2014.igem.org

(Difference between revisions)
(Created page with "> {{:Team:British_Columbia/Templates/MainHeader}} <html> <div class="container"> <h1>Liquid Media</h1> <h2>Supplies:</h2> <ul> <li>dH2O</li> <li>Autoclavable container (500mL...")
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<h2>Steps:</h2>
<h2>Steps:</h2>
<ol>
<ol>
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1. Pour appropriate amount of diH2O (400mL) into autoclavable container.
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<li> Pour appropriate amount of diH2O (400mL) into autoclavable container.</li>
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2. Add LB broth powder (Check amount on stock container).
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<li> Add LB broth powder (Check amount on stock container).</li>
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3. Mix as well as you can by swirling and gentle shaking.
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<li> Mix as well as you can by swirling and gentle shaking.</li>
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4. Autoclave.
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<li> Autoclave:</li>
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<br>
 +
<ol type="a">
 +
<li>Mark bottle with autoclave tape, make sure the lid is loose</li>
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<li>Follow directions on autoclave</li>
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*Mark bottle with autoclave tape, make sure the lid is loose*
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-
*Follow directions on autoclave*
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<li>Set to 30 min, turn timer counterclockwise, check back later.</li>
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*Set to 30 min, turn timer counterclockwise* (CHECK LATER)
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<li>Before removing from autoclave make sure the pressure is at 0</li>
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*Before removing from autoclave make sure the pressure is at 0*
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<li>Use heat-resistant gloves to remove from Autoclave</li>
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*Use heat-resistant gloves to remove from Autoclave*
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</ol type="a">
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<li> Cool until you can hold your wrist against the bottle for 5 seconds without pain.</li>
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5. Cool until you can hold your wrist against the bottle for 5 seconds without pain.
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<li> Add antibiotics (final concentrations: A-100-150μg/mL, K-50μg/mL, C-25μg/mL, Tet-10-15μg/mL); if using recommended stock solution concentrations, add 1μL of antibiotic stock per 1mL of media.</li>
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6. Add antibiotics (final concentrations: A-100-150μg/mL, K-50μg/mL, C-25μg/mL, Tet-10-15μg/mL); if using recommended stock solution concentrations, add 1μL of antibiotic stock per 1mL of media.
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<li> Swirl gently to mix antibiotics.</li>
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7. Swirl gently to mix antibiotics.
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<li> For storage, seal with parafilm and label according to the colour scheme. Keep at 4°C. If
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using a light-sensitive antibiotic, wrap in foil as well.</li>
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8. For storage, seal with parafilm and label according to the colour scheme. Keep at 4°C. If
+
-
using a light-sensitive antibiotic, wrap in foil as well.
+
</ul>
</ul>

Revision as of 00:25, 18 October 2014

> 2014 UBC iGEM

Liquid Media

Supplies:

  • dH2O
  • Autoclavable container (500mL Glass Bottles)
  • Antibiotics (Stock solutions: Amp - 100mg/mL in 50% ethanol, Kan - 50mg/mL in H2O, Chl - 35mg/mL in 100%? ethanol, Tet - 15 mg/ml in 50% ethanol - all at -20°C)
  • Tinfoil (if using Tet)
  • LB broth powder
  • Colour scheme: Amp - red, Kan - blue, Chl - black, Tet - green

Steps:

  1. Pour appropriate amount of diH2O (400mL) into autoclavable container.
  2. Add LB broth powder (Check amount on stock container).
  3. Mix as well as you can by swirling and gentle shaking.
  4. Autoclave:

    1. Mark bottle with autoclave tape, make sure the lid is loose
    2. Follow directions on autoclave
    3. Set to 30 min, turn timer counterclockwise, check back later.
    4. Before removing from autoclave make sure the pressure is at 0
    5. Use heat-resistant gloves to remove from Autoclave
  5. Cool until you can hold your wrist against the bottle for 5 seconds without pain.
  6. Add antibiotics (final concentrations: A-100-150μg/mL, K-50μg/mL, C-25μg/mL, Tet-10-15μg/mL); if using recommended stock solution concentrations, add 1μL of antibiotic stock per 1mL of media.
  7. Swirl gently to mix antibiotics.
  8. For storage, seal with parafilm and label according to the colour scheme. Keep at 4°C. If using a light-sensitive antibiotic, wrap in foil as well.

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