Team:Warsaw/Calendar-Main/11 August 2008

From 2008.igem.org

(Difference between revisions)
Line 23: Line 23:
[photo of the gel is to be placed here]
[photo of the gel is to be placed here]
 +
<h3>Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>)</h3>
 +
<h4>Piotr</h4>
 +
<p>
 +
Test was conducted in <i>E.coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosetta</a> strain expressing omp_omega_A_alfa (with and without induction) and omp_A_alfa.
 +
<ol>
 +
<li>Spinnign</li>
 +
<li>Suspending</li>
 +
<li>Adding of lysis buffer</li>
 +
<li>Boiling</li>
 +
<li>Putting into poliacrylamide gel</li>
 +
<li>Transfer onto nitrocellulose</li>
 +
<li>Blocking</li>
 +
<li>Anti-A antibody binding</li>
 +
<li>Washing</li>
 +
<li>Anti-rabbit antibody binding</li>
 +
<li>Developing with BCIP and NBT</li>
 +
</ol>
 +
</p>
 +
 +
 +
[photo of the gel is to be placed here]
 +
<p>We didn't observe differences in espression and degradation in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#rosetta">Rosettas</a> nor in <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>. Therefore we suppose that degradation of the fusions is caused by other factor than Lon and OmpT proteases.</p>

Revision as of 13:33, 12 October 2008

Gallery Bricks Notebook Team Project Home


Previous day
return to main notebook page
Previous entry
Interactive list of topics
next notebook entry

 




Checking for presence of A on the cell membrane
Piotr

  1. Pouring bacteria by drops onto nitrocellulose
  2. Drying
  3. Blocking
  4. Anti-A antibody binding
  5. Washing
  6. Anti-rabbit antibody binding
  7. Developing with BCIP and NBT

[photo of the gel is to be placed here]

Checking whether degradation of the fusions with OmpA is caused by Lon and OmpT proteases (present in TOP10)

Piotr

Test was conducted in E.coli Rosetta strain expressing omp_omega_A_alfa (with and without induction) and omp_A_alfa.

  1. Spinnign
  2. Suspending
  3. Adding of lysis buffer
  4. Boiling
  5. Putting into poliacrylamide gel
  6. Transfer onto nitrocellulose
  7. Blocking
  8. Anti-A antibody binding
  9. Washing
  10. Anti-rabbit antibody binding
  11. Developing with BCIP and NBT

[photo of the gel is to be placed here]

We didn't observe differences in espression and degradation in Rosettas nor in TOP10. Therefore we suppose that degradation of the fusions is caused by other factor than Lon and OmpT proteases.

Retrieved from "http://2008.igem.org/Team:Warsaw/Calendar-Main/11_August_2008"