Team:Warsaw/Calendar-Main/3 September 2008

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(New page: {{WarNotebook}} <!-- do not edit above me! --> Oczyszczanie białek: A-alfa, Z-alfa i Z-omega Piotr zawieszenie próbek w PBS, sonikacja, zwirowanie, do supernatantu i osadu osobn...)
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<h3>Purification of proteins:  A-alpha, Z-alpha and Z-omega<br>Piotr</h3>
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<p><ol><li>Samples were resuspended in PBS, sonicated and centrifuged.</li>
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<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>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)</p>
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Oczyszczanie białek: A-alfa, Z-alfa i Z-omega
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Piotr
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zawieszenie próbek w PBS, sonikacja, zwirowanie, do supernatantu i osadu osobno dodanie buforu lizującego (opisać skład gdzieś) gotować przez 10 minut, na żel poliakryl 12% (zrobić protokół) nałożyć  ilość zlizowanych bakterii odpowiadającą 100mikrolitrom chodowli o OD=1, elektroforeza (zrobić protokół), barwienie i odbarwianie qmasim (zrobić protokuł)
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wybranie optymalnych warunków
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zaszczepienie A-alfa, Z-alfa i Z-omega
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Revision as of 22:33, 5 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)

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