Team:Warsaw/Calendar-Main/29 May 2008

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<h3>Rifampicin test #3</h3>
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<h4>Michał L., Ewa, Marcin, Piotr</h4>
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<p><ol>
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<li> Added 50 μL <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  carrying <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5+AID</a> (in liquid LB) into liquid LB.</li>
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<li> Added 50 μL <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  carrying <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5+AID</a> (in liquid LB) into liquid LB + 0.02% arabinose.</li>
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<li> Added 50 μL <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  carrying <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A+AID</a> (in liquid LB) into liquid LB.</li>
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1. Add 50 μL E.coli TOP10 carrying pMPM-T5+AID (in liquid LB) into liquid LB
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<li> Added 50 μL <i>E. coli</i> <a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#top10">TOP10</a>  carrying <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A+AID</a> (in liquid LB) into liquid LB + 1 mM IPTG.</li>
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2. Add 50 μL E.coli TOP10 carrying pMPM-T5+AID (in liquid LB) into liquid LB + 0.02% arabinose
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<li> Cultures were incubated for 5 hours.</li>
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3. Add 50 μL E.coli TOP10 carrying pTrc99A+AID (in liquid LB) into liquid LB
 
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4. Add 50 μL E.coli TOP10 carrying pTrc99A+AID (in liquid LB) into liquid LB + 1mM IPTG
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<li> 1 ml (2x) from p.1, p.2, p.3, p.4 were plated on LB + rifampicin. </li>
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5. Wait for 5 hours
 
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6. OD measurment
 
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7. 1 ml (2x) from p.1, p.2, p.3, p.4 (LB + riphampicin)
 
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<h3>PCR results of translational fusion for the A+AT construct.<br> Lane 1 - 25 cycles, lane 2 - 20 cycles.</h3>
 
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https://static.igem.org/mediawiki/2008/0/08/Tranlacyjna_pcr_29_05_2008.jpg
 
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<li>OD measurement of above cultures.</li>
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</ol></p>
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</html>
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Rifampicin test #3

Michał L., Ewa, Marcin, Piotr

  1. Added 50 μL E. coli TOP10 carrying pMPM-T5+AID (in liquid LB) into liquid LB.
  2. Added 50 μL E. coli TOP10 carrying pMPM-T5+AID (in liquid LB) into liquid LB + 0.02% arabinose.
  3. Added 50 μL E. coli TOP10 carrying pTrc99A+AID (in liquid LB) into liquid LB.
  4. Added 50 μL E. coli TOP10 carrying pTrc99A+AID (in liquid LB) into liquid LB + 1 mM IPTG.
  5. Cultures were incubated for 5 hours.
  6. 1 ml (2x) from p.1, p.2, p.3, p.4 were plated on LB + rifampicin.
  7. OD measurement of above cultures.

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