Team:Bielefeld-CeBiTec/Notebook/Media

From 2014.igem.org

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<h1>Media</h1>
<h1>Media</h1>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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        <div class="tab" id="LBmedium">
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            <div class="show">
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                <a href="#LBmedium">LB medium</a>
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<a href="https://static.igem.org/mediawiki/2014/b/bb/Bielefeld-CeBiTec_2014-08-12_LB-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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            </div>
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            <div class="hide">
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                <a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6>LB medium  <a href="https://static.igem.org/mediawiki/2014/b/bb/Bielefeld-CeBiTec_2014-08-12_LB-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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            </div>
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            <div class="content">
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<ul class="protocol">
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<li> 10 g Trypton </li>
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<li> 5 g yeast extract </li>
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<li> 10 g NaCl </li>
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<li> 12 g Agar-Agar (for plates) </li>
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            </div>
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        </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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        <div class="tab" id="M9medium">
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            <div class="show">
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                <a href="#M9medium">M9 medium</a>
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<a href="https://static.igem.org/mediawiki/2014/8/85/Bielefeld-CeBiTec_2014-08-12_M9-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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            </div>
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            <div class="hide">
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                <a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6>M9 medium <a href="https://static.igem.org/mediawiki/2014/8/85/Bielefeld-CeBiTec_2014-08-12_M9-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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            </div>
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            <div class="content">
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                <ul style="margin-left:10%; margin-right:10%" type="disc">
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                <li>To prepare 1 liter of M9 minimal medium add the following components to 836.7 mL sterile distilled  water. To avoid precipitation, start with CaCl2 and mix the solution thoroughly after addition of a  component. </li>
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                <li> 100 mL 10 X M9 salt solution </li>
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                <li> 50 mL 20 X carbon source </li>
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                <li> 1 mL 1000 X trace element solution </li>
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                <li> 1 mL autoclaved 1 M MgSO4 </li>
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                <li> 0.3 mL autoclaved 1 M CaCl2 </li>
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                <li> 1 mL filter sterilized 1 g/L biotin </li>
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                <li> 1 mL filter sterilized 1 g/L thiamin </li> </ul>
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            </div>
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        </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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        <div class="tab" id="M9saltstocksolution">
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            <div class="show">
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                <a href="#M9saltstocksolution">M9 salt stock solution </a>
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<a href="https://static.igem.org/mediawiki/2014/1/1a/Bielefeld-CeBiTec_2014-08-12_M9-salt-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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            </div>
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            <div class="hide">
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                <a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6>M9 salt stock solution <a href="https://static.igem.org/mediawiki/2014/1/1a/Bielefeld-CeBiTec_2014-08-12_M9-salt-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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            </div>
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            <div class="content">
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                <ul style="margin-left:10%; margin-right:10%" type="disc">
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                <li> 75.2 g Na2HPO4 x 2H2O </li>
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                <li> 30 g KH2PO4 </li>
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                <li> 5 g NaCl </li>
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                <li> 5 g NH4Cl </li>
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                <li> Dissolve the salts in 800 mL water and adjust the pH to 7.2 with NaOH. Add water to a final volume of 1 L and autoclave for 15 minutes at 121 °C. </li></ul>
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          </div>
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      </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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    <div class="tab" id="SOC-medium">
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      <div class="show">
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        <a href="#SOC-medium"> SOC-medium </a>
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<a href="https://static.igem.org/mediawiki/2014/5/53/Bielefeld-CeBiTec_2014-08-12_SOC-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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      </div>
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      <div class="hide">
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        <a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6>SOC-medium <a href="https://static.igem.org/mediawiki/2014/5/53/Bielefeld-CeBiTec_2014-08-12_SOC-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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      </div>
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      <div class="content">
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        <ul style="margin-left:10%; margin-right:10%" type="disc">
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          <li> Add the following components for 900 ml of distilled H2O: </li>
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          <ul> <li>20 g Trypton </li>
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              <li> 5 g Bacto Yeast Extract </li>
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              <li> 2 mL of 5 M NaCl </li>
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              <li> 2.5 ml of 1 M KCl </li>
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              <li> 10 ml of 1 M MgCl2 </li>
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              <li> 10 ml of 1 M MgSO4 </li>
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              <li> 20 ml of 1 M glucose </li> </ul> </ul>
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      </div>
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    </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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    <div class="tab" id="50XTAEStockSolution">
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      <div class="show">
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        <a href="#50XTAEStockSolution"> 50X TAE Stock Solution </a>
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<a href="https://static.igem.org/mediawiki/2014/e/ed/Bielefeld-CeBiTec_2014-08-12_50x-TAE-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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      </div>
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      <div class="hide">
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        <a  style="font-size:24px" href="#"><p style="margin-left:30%"> <h6>50X TAE Stock Solution <a href="https://static.igem.org/mediawiki/2014/e/ed/Bielefeld-CeBiTec_2014-08-12_50x-TAE-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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      </div>
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      <div class="content">
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        <ul style="margin-left:10%; margin-right:10%" type="disc">
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          <li> For each litre of solution: </li>
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          <ul> <li> 242 g Tris Base (MW=121.1) </li>
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              <li> 57.1 mL Glacial Acetic Acid </li>
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              <li> 100 mL 0.5 M EDTA </li> </ul> <br>
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          <li> mix Tris with stir bar to dissolve in about 600 mL of ddH2O. </li>
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          <li> add the EDTA and Acetic Acid. </li>
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          <li> bring final volume to 1 L with ddH20. </li>
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          <li> store at room temperature. </li> <br>
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          <li> Note: Final (1x) working concentration: </li>
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          <ul> <li> 0.04 M Tris - Acetate </li>
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              <li> 0.001 M EDTA </li> </ul> </ul>
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      </div>
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    </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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    <div class="tab" id="50XModifiedTAEStockSolution">
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      <div class="show">
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        <a href="#50XModifiedTAEStockSolution"> 50X Modified TAE Stock Solution </a>
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<a href="https://static.igem.org/mediawiki/2014/3/3b/Bielefeld-CeBiTec_2014-08-12_50x-modified-TAE-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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      </div>
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      <div class="hide">
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        <a  style="font-size:24px" href="#"><p style="margin-left:30%"> <h6>50X Modified TAE Stock Solution <a href="https://static.igem.org/mediawiki/2014/3/3b/Bielefeld-CeBiTec_2014-08-12_50x-modified-TAE-stock-solution.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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      </div>
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      <div class="content">
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        <ul style="margin-left:10%; margin-right:10%" type="disc">
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          <li> For each litre of solution: </li>
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          <ul> <li> 242 g Tris Base (MW=121.1) </li>
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              <li> 57.1 mL Glacial Acetic Acid </li>
 +
              <li> 10 mL 0.5 M EDTA </li> </ul> <br>
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          <li> mix Tris with stir bar to dissolve in about 600 mL of ddH2O. </li>
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          <li> add the EDTA and Acetic  Acid, pH to 8.0. </li>
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          <li> bring final volume to 1 L with ddH2O. </li>
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          <li> store at room temperature. </li> <br>
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 +
          <li> Note: Final (1x) working concentration: </li>
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          <ul> <li> 0.04 M Tris - Acetate </li>
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              <li> 0.0001 M EDTA </li> </ul> </ul>
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      </div>
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    </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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  <div id="text">
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<div class="tab" id="TAEbuffer">
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            <div class="show">
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                <a href="#TAEbuffer">TAE buffer</a>
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<a href="https://static.igem.org/mediawiki/2014/0/0e/Bielefeld-CeBiTec_2014-08-12_TAE-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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            </div>
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            <div class="hide">
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                <a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6>TAE buffer <a href="https://static.igem.org/mediawiki/2014/0/0e/Bielefeld-CeBiTec_2014-08-12_TAE-medium.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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            </div>
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            <div class="content">
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                <ul style="margin-left:10%; margin-right:10%" type="disc">
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<li> 1 L of 50x TAE buffer </li>
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<li> 242.48 g Tris </li>
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<li> 41.02 g sodium acetate </li>
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<li> 18.612 g EDTA </li>
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<li> Adjust pH to 7.8 with acetic acid </li>
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<li> Solve in dH2O </li>
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<li> Dilute 20 mL 50x stock in 1L dH2O for 1x Buffer for PAGE</li> </ul>
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            </div>
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        </div>
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  </div>
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</div>
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<div class="element" style="margin:10px 10px 10px 10px; padding:10px 10px 10px 10px">
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<div id="text">
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<div class="tab" id="Loadingbuffer">
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<div class="show">
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<a href="#Loadingbuffer"> Loading buffer </a>
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<a href="https://static.igem.org/mediawiki/2014/.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a>
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</div>
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<div class="hide">
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<a  style="font-size:24px" href="#"><p style="margin-left:30%"><h6> Loading Buffer <a href="https://static.igem.org/mediawiki/2014/.pdf"> <img src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png" height="30px"> </a></h6></p></a>
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</div>
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<div class="content">
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<ul style="margin-left:10%; margin-right:10%" type="disc">
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<li>292.243g/mol 1mM EDTA </li>
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<li> 0.025g 0.05% (w/v) BPB </li>
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<li> 0.025g 0.05% (w/v) Xylene Cyanol </li>
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<li> Solve in H2O </li>
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<li> Adjust color to green with HCl </li>
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<li> Dilute with glycerol to 50:50 </li> </ul>
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</div>
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</div>
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</div>
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</div>
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Revision as of 13:53, 12 August 2014


Media

LB medium

LB medium

  • 10 g Trypton
  • 5 g yeast extract
  • 10 g NaCl
  • 12 g Agar-Agar (for plates)
M9 medium

M9 medium

  • To prepare 1 liter of M9 minimal medium add the following components to 836.7 mL sterile distilled water. To avoid precipitation, start with CaCl2 and mix the solution thoroughly after addition of a component.
  • 100 mL 10 X M9 salt solution
  • 50 mL 20 X carbon source
  • 1 mL 1000 X trace element solution
  • 1 mL autoclaved 1 M MgSO4
  • 0.3 mL autoclaved 1 M CaCl2
  • 1 mL filter sterilized 1 g/L biotin
  • 1 mL filter sterilized 1 g/L thiamin
M9 salt stock solution

M9 salt stock solution

  • 75.2 g Na2HPO4 x 2H2O
  • 30 g KH2PO4
  • 5 g NaCl
  • 5 g NH4Cl
  • Dissolve the salts in 800 mL water and adjust the pH to 7.2 with NaOH. Add water to a final volume of 1 L and autoclave for 15 minutes at 121 °C.
SOC-medium

SOC-medium

  • Add the following components for 900 ml of distilled H2O:
    • 20 g Trypton
    • 5 g Bacto Yeast Extract
    • 2 mL of 5 M NaCl
    • 2.5 ml of 1 M KCl
    • 10 ml of 1 M MgCl2
    • 10 ml of 1 M MgSO4
    • 20 ml of 1 M glucose
50X TAE Stock Solution

50X TAE Stock Solution

  • For each litre of solution:
    • 242 g Tris Base (MW=121.1)
    • 57.1 mL Glacial Acetic Acid
    • 100 mL 0.5 M EDTA

  • mix Tris with stir bar to dissolve in about 600 mL of ddH2O.
  • add the EDTA and Acetic Acid.
  • bring final volume to 1 L with ddH20.
  • store at room temperature.

  • Note: Final (1x) working concentration:
    • 0.04 M Tris - Acetate
    • 0.001 M EDTA
50X Modified TAE Stock Solution

50X Modified TAE Stock Solution

  • For each litre of solution:
    • 242 g Tris Base (MW=121.1)
    • 57.1 mL Glacial Acetic Acid
    • 10 mL 0.5 M EDTA

  • mix Tris with stir bar to dissolve in about 600 mL of ddH2O.
  • add the EDTA and Acetic Acid, pH to 8.0.
  • bring final volume to 1 L with ddH2O.
  • store at room temperature.

  • Note: Final (1x) working concentration:
    • 0.04 M Tris - Acetate
    • 0.0001 M EDTA
TAE buffer

TAE buffer

  • 1 L of 50x TAE buffer
  • 242.48 g Tris
  • 41.02 g sodium acetate
  • 18.612 g EDTA
  • Adjust pH to 7.8 with acetic acid
  • Solve in dH2O
  • Dilute 20 mL 50x stock in 1L dH2O for 1x Buffer for PAGE
Loading buffer

Loading Buffer

  • 292.243g/mol 1mM EDTA
  • 0.025g 0.05% (w/v) BPB
  • 0.025g 0.05% (w/v) Xylene Cyanol
  • Solve in H2O
  • Adjust color to green with HCl
  • Dilute with glycerol to 50:50

Retrieved from "http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Media"