Team:Grinnell/Recipes

From 2011.igem.org

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<h1>Recipes</h1>
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<h2><a name="LB"></a>L Broth</h2>
<h2><a name="LB"></a>L Broth</h2>
For 1L:<ul>
For 1L:<ul>
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<li>5g Yeast extract</li>
<li>5g Yeast extract</li>
<li>5g NaCl</li></ul>
<li>5g NaCl</li></ul>
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<h2><a name="PYE"></a>PYE Broth</h2>
<h2><a name="PYE"></a>PYE Broth</h2>
For 1L:<ul>
For 1L:<ul>
<li>2g Bacto peptone</li>
<li>2g Bacto peptone</li>
<li>1g Yeast extract</li>
<li>1g Yeast extract</li>
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<li>1mL 1M MgSO4&sdot;7H2O</li>
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<li>1mL 1M MgSO<sub>4</sub>&sdot;7H<sub>2</sub>O</li>
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<li>1mL 0.5M CaCl2</li></ul>
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<li>1mL 0.5M CaCl<sub>2</sub></li></ul>
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<h2><a name="M2G"></a>M2G Minimal Broth</h2>
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<ol>
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<li>For 1L of 20x M2 Salts:<ul>
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<li>17.4g Na<sub>2</sub>HPO<sub>4</sub></li>
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<li>10.6g KH<sub>2</sub>PO<sub>4</sub></li>
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<li>10g NH<sub>4</sub>Cl</li>
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</ul></li>
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<li>Autoclave this sol'n.</li>
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<li>To make M2G:
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<ol>
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<li>Dilute 20x M2 salts sol'n to 1x and autoclave.</li>
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<li>Add the following sterile sol'ns:
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<ul>
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<li>1mL 0.5M MgSo<sub>4</sub></li>
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<li>10mL 20% Glucose</li>
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<li>1mL 10mM FeSO<sub>4</sub>, 10mM EDTA (Sigma #F-0518, kept at 4&deg;C)</li>
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<li>500&mu;L 1M CaCl<sub>2</sub> (Note: add this last to avoid precipitation)</li>
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</ul>
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</li>
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</ol>
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</li>
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</ol>
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<h2><a name="Agar"></a>Agar Plates</h2>
<h2><a name="Agar"></a>Agar Plates</h2>
Add 15g Bacto-agar to 1L of liquid media before autoclaving.
Add 15g Bacto-agar to 1L of liquid media before autoclaving.
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<h2><a name="dGel"></a>0.7% Agarose Gel</h2>
 
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<h2><a name="Anti"></a>Antibiotics Kitchen</h2>
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<table>
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<tr>
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<th>Antibiotic</th>
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<th>Color Code</th>
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<th>Stock (mg/mL)</th>
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<th>LB Plate Conc (&mu;g/mL)</th>
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<th>PYE Plate Conc (&mu;g/mL)</th>
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</tr>
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<tr>
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<td style="text-align:left">Ampicillin (amp)</td>
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<td>Red</td>
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<td>50</td>
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<td>100</td>
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<td>50</td>
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</tr>
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<tr>
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<td style="text-align:left">Carbenicillin (carb)</td>
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<td>Red</td>
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<td>50</td>
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<td>50</td>
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<td>50</td>
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</tr>
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<tr>
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<td style="text-align:left">Chloramphenicol (chlor)</td>
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<td>2 Orange</td>
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<td>5 (in EtOH)</td>
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<td>30</td>
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<td>1</td>
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</tr>
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<tr>
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<td style="text-align:left">Kanamycin (kan)</td>
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<td>blue</td>
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<td>50</td>
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<td>50</td>
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<td>25</td>
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</tr>
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<tr>
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<td style="text-align:left">Nalidixic acid (nal)</td>
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<td>Green</td>
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<td>10</td>
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<td>20</td>
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<td>20</td>
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</tr>
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<tr>
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<td style="text-align:left">Tetracycline (tet)</td>
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<td>Black</td>
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<td>2 (in EtOH)</td>
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<td>12</td>
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<td>&ndash;</td>
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</tr>
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<tr>
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<td style="text-align:left">Oxytetracycline (oxytet)</td>
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<td>Black</td>
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<td>2 (in EtOH)</td>
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<td>&minus;</td>
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<td>2</td>
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</tr>
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<tr>
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<td style="text-align:left">Spectinomycin (spec)</td>
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<td>Purple</td>
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<td>20</td>
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<td>50</td>
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<td>100</td>
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</tr>
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</table>
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<br/><b>Note</b>: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures).
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<h2><a name="freezerStocks"></a>Freezer Stocks</h2>
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To an appropriate tube add:
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<ul>
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    <li>700&mu;L overnight liquid culture</li>
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    <li>300&mu;L 50% glycerol sol'n</li>
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</ul>
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Then place in storage at -80&deg;C.
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Latest revision as of 19:55, 28 September 2011

Grinnell Menubar

Recipes

L Broth

For 1L:
  • 10g Tryptone
  • 5g Yeast extract
  • 5g NaCl

PYE Broth

For 1L:
  • 2g Bacto peptone
  • 1g Yeast extract
  • 1mL 1M MgSO4⋅7H2O
  • 1mL 0.5M CaCl2

M2G Minimal Broth

  1. For 1L of 20x M2 Salts:
    • 17.4g Na2HPO4
    • 10.6g KH2PO4
    • 10g NH4Cl
  2. Autoclave this sol'n.
  3. To make M2G:
    1. Dilute 20x M2 salts sol'n to 1x and autoclave.
    2. Add the following sterile sol'ns:
      • 1mL 0.5M MgSo4
      • 10mL 20% Glucose
      • 1mL 10mM FeSO4, 10mM EDTA (Sigma #F-0518, kept at 4°C)
      • 500μL 1M CaCl2 (Note: add this last to avoid precipitation)

Agar Plates

Add 15g Bacto-agar to 1L of liquid media before autoclaving.

Antibiotics Kitchen

Antibiotic Color Code Stock (mg/mL) LB Plate Conc (μg/mL) PYE Plate Conc (μg/mL)
Ampicillin (amp) Red 50 100 50
Carbenicillin (carb) Red 50 50 50
Chloramphenicol (chlor) 2 Orange 5 (in EtOH) 30 1
Kanamycin (kan) blue 50 50 25
Nalidixic acid (nal) Green 10 20 20
Tetracycline (tet) Black 2 (in EtOH) 12 –
Oxytetracycline (oxytet) Black 2 (in EtOH) − 2
Spectinomycin (spec) Purple 20 50 100

Note: for liquid cultures in PYE broth, use half the concentration that would be used for a PYE plate (LB remains the same for liquid and sold cultures).

Freezer Stocks

To an appropriate tube add:
  • 700μL overnight liquid culture
  • 300μL 50% glycerol sol'n
Then place in storage at -80°C.

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