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Team:Warsaw/Calendar-Main/26 June 2008
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| - | <h3>Change of the reporter from | + | <h3>Change of the reporter from <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC320</a> with B-galactosidase to GFP or RFP</h3> |
<h4>Piotr, Weronika</h4> | <h4>Piotr, Weronika</h4> | ||
<ol> | <ol> | ||
| - | <li>Isolation of | + | <li>Isolation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pZC320>pZC30</a>.</li> |
| - | <li>Inoculation of E.coli TOP10 carrying | + | <li>Inoculation of E.coli TOP10 carrying <a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A2>pSB1A2</a> standard plasmids carrying parts: <A href=http://partsregistry.org/wiki/index.php?title=Part:BBa_E0840>BBa_E0840</a>(GFP genetrator) and <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_J04450>BBa_J04450</a> (RFP generator). |
</li></ol> | </li></ol> | ||
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| - | <h3>Preparation of constructs | + | <h3>Preparation of constructs: OmpA_alpha and OmpA_omega</h3><h4>Michał K.</h4> |
<p>Repetition of PCRs, gel-out isolations and digests from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_June_2008">17 June.</a></p> | <p>Repetition of PCRs, gel-out isolations and digests from <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/17_June_2008">17 June.</a></p> | ||
| - | <h3><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcl">Polymerase Chain Ligation (PCL)</a></ | + | <h3>Preparation of alpha-A and omega-A fusions</h3><h4>Michał L., Ewa, Marcin</h4> |
| + | <b><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcl">Polymerase Chain Ligation (PCL)</a></b> | ||
<table id="result"> | <table id="result"> | ||
<tr><th>Product</th><th>Templates</th><th>Primers</th><th>Product length</th></tr> | <tr><th>Product</th><th>Templates</th><th>Primers</th><th>Product length</th></tr> | ||
| - | <tr><th>Alpha-A fusion</th><td>Alpha-linker + linker-A</td><td>AlphaL+SacI + AP+NotI </td><td> | + | <tr><th>Alpha-A fusion</th><td>Alpha-linker + linker-A</td><td>AlphaL+SacI + AP+NotI </td><td>1100 bp</td></tr> |
| - | <tr><th>Omega-A fusion</th><td>Omega-linker + linker-A</td><td>OmegaL+SacI + AP+NotI </td><td> | + | <tr><th>Omega-A fusion</th><td>Omega-linker + linker-A</td><td>OmegaL+SacI + AP+NotI </td><td>750 bp</td></tr> |
</table> | </table> | ||
<br><br> | <br><br> | ||
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<tr><td>94°C</td><td>0:30</td><td rowspan="3">28 cycles</td></tr> | <tr><td>94°C</td><td>0:30</td><td rowspan="3">28 cycles</td></tr> | ||
<tr><td>48°C-62°C</td><td>0:45</td></tr> | <tr><td>48°C-62°C</td><td>0:45</td></tr> | ||
| - | <tr><td>68°C</td><td> | + | <tr><td>68°C</td><td>1:30</td></tr> |
<tr><td>68°C</td><td>10:00</td></tr> | <tr><td>68°C</td><td>10:00</td></tr> | ||
<tr><td>4°C</td><td>infinite</td></tr> | <tr><td>4°C</td><td>infinite</td></tr> | ||
Latest revision as of 14:02, 28 October 2008
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Change of the reporter from pZC320 with B-galactosidase to GFP or RFPPiotr, Weronika
Preparation of constructs: OmpA_alpha and OmpA_omegaMichał K.Repetition of PCRs, gel-out isolations and digests from 17 June. Preparation of alpha-A and omega-A fusionsMichał L., Ewa, MarcinPolymerase Chain Ligation (PCL)
PCL program for fusion A with alpha and omega
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