Team:UC Davis/Notebook/Protocols

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<li>8. Resuspend with 300 uL of B-PER Protein Extraction Reagent and follow the instructions provided with that kit. After this, there should be samples of culture media, and soluble and insoluble whole cell lysate.</li>
<li>8. Resuspend with 300 uL of B-PER Protein Extraction Reagent and follow the instructions provided with that kit. After this, there should be samples of culture media, and soluble and insoluble whole cell lysate.</li>
<li>9. Take 15 uL of each sample, including controls, and run on western blot.</li>
<li>9. Take 15 uL of each sample, including controls, and run on western blot.</li>
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    </div>
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<p class="menu_head">pNPB Assay</p>
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    <div class="menu_body">
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<p>Materials</p>
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<ul>
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<li>pNPB buffer (should be made and used in fumehood due to acetonitriles toxicity)
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<ul><li>10mM pNPB in acetonitrile</li>
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<li>1:4:95 ratio of acetonitrile pNPB solution, 100% ethanol and 50mM Tris-HCL (pH 8)</li></ul></li>
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<li>LB with specific resistance</li>
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<li>Cell Culture</li>
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<li>96 well plate</li></ul>
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<br>
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<p>Protocol</p>
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<ul><li>First assign each well of the plate except for outer wells which should be filled with LB to prevent evaporation of medium.</li>
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<li>Fill wells with 95 uL of LB</li>
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<li>Fill wells with 5 uL of cell culture</li>
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<li>In fumehood, add 100 uL of bufer to each well</li>
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<li>Run in Tecan taking both ODs and absorbance 405 readings</li>
     </div>
     </div>

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