Team:Warsaw/Calendar-Main/3 September 2008

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<h3>Purification of proteins:  A-alpha, Z-alpha and Z-omega<br>Piotr</h3>
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<h3>Purification of proteins:  A-alpha, Z-alpha and Z-omega</h3><h4>Piotr</h4>
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<p><ol><li>Samples were resuspended in PBS, sonicated and centrifuged.</li>  
<p><ol><li>Samples were resuspended in PBS, sonicated and centrifuged.</li>  
<li>Lysis buffer were added separately to pellet and supernatant, then samples were boiled for 10 minutes. </li>
<li>Lysis buffer were added separately to pellet and supernatant, then samples were boiled for 10 minutes. </li>
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<li>Lysates loaded on 12% poliacrylamide gel (amount relating to 100 ul of OD=1,0 culture)</li>
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<li>Lysates loaded on 12% poliacrylamide gel (amount relating to 100 ul of OD=1,0 culture).</li>
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<li>Gel stained with Coomassie Blue. Optimal induction conditions chosen</li>
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<li>Gel stained with Coomassie Blue. Optimal induction conditions chosen.</li>
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<li>A-alpha, Z-alpha and Z-omega inoculated in the overnight culture (Rosetta strain)</li></ol></p>
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<li>A-alpha, Z-alpha and Z-omega inoculated in the overnight culture (Rosetta strain).</li></ol></p>
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Revision as of 22:41, 12 October 2008

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Purification of proteins: A-alpha, Z-alpha and Z-omega

Piotr

  1. Samples were resuspended in PBS, sonicated and centrifuged.
  2. Lysis buffer were added separately to pellet and supernatant, then samples were boiled for 10 minutes.
  3. Lysates loaded on 12% poliacrylamide gel (amount relating to 100 ul of OD=1,0 culture).
  4. Gel stained with Coomassie Blue. Optimal induction conditions chosen.
  5. A-alpha, Z-alpha and Z-omega inoculated in the overnight culture (Rosetta strain).