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| Line 28: |
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| | </p> | | </p> |
| - | <div class="center"><div class="thumb tnone"><div class="thumbinner" style="width:302px;"><a href="/Image:Unipv_digestion_J231xx_I6.jpg" class="image" title="J231xx and I6 digestion"><img alt="J231xx and I6 digestion" src="https://static.igem.org/mediawiki/2010/thumb/c/c8/Unipv_digestion_J231xx_I6.jpg/300px-Unipv_digestion_J231xx_I6.jpg" width="300" height="142" border="0" class="thumbimage" /></a> <div class="thumbcaption"><div class="magnify"><a href="/Image:Unipv_digestion_J231xx_I6.jpg" class="internal" title="Ingrandisci"><img src="/wiki/skins/common/images/magnify-clip.png" width="15" height="11" alt="" /></a></div>J231xx and I6 digestion</div></div></div></div>
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| - | <p>After MiniPrep, purified DNA was quantified with NanoDrop.
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| - | </p>
| |
| - | <table border="1" align="center">
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| - | <tr>
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| - | <td> I6 </td><td> 193 ng/ul
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| - | </td></tr>
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| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23118'>BBa_J23118</A> </td><td> 107,9 ng/ul
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| - |
| |
| - | </td></tr>
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| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23110'>BBa_J23110</A> </td><td> 83 ng/ul
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| - | </td></tr>
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| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23114'>BBa_J23114</A> </td><td> 89,8 ng/ul
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| - | </td></tr>
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| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23116'>BBa_J23116</A> </td><td> 82,7 ng/ul
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| - |
| |
| - | </td></tr></table>
| |
| - | <p><br />
| |
| - | Digestion of:
| |
| - | </p>
| |
| - | <table border="1" align="center">
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| - | <tr>
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| - | <td> <i>Culture</i> </td><td> <i>Kind</i> </td><td> <i>Final reaction volume (ul) </i> </td><td> <i>DNA (ul)</i> </td><td> <i>H20 (ul)</i> </td><td> <i>Enzyme 1</i> </td><td> <i>Enzyme 2</i> </td><td> <i>Buffer H</i>
| |
| - |
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td> I6 </td><td> Insert </td><td> 25 </td><td> 9,3 </td><td> 11,2 </td><td> 1 XbaI </td><td> 1 PstI </td><td> 2,5
| |
| - |
| |
| - | </td></tr>
| |
| - | <tr>
| |
| - | <td> I6-bis </td><td> Insert </td><td> 25 </td><td>9,3 </td><td> 11,2 </td><td> 1 XbaI </td><td> 1 PstI </td><td> 2,5
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| - |
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23118'>BBa_J23118</A> </td><td> Vector </td><td> 25 </td><td> 9,3 </td><td> 11,2 </td><td> 1 SpeI </td><td> 1 PstI </td><td> 2,5
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| - |
| |
| - | </td></tr>
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| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23110'>BBa_J23110</A> </td><td> Vector </td><td> 25 </td><td>12 </td><td> 8,5 </td><td> 1 SpeI </td><td> 1 PstI </td><td> 2,5
| |
| - |
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23114'>BBa_J23114</A> </td><td> Vector </td><td> 25 </td><td> 11,2 </td><td> 9,3 </td><td> 1 SpeI </td><td> 1 PstI </td><td> 2,5
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| - |
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td> <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23116'>BBa_J23116</A> </td><td> Vector </td><td> 25 </td><td> 12,1 </td><td> 8,4 </td><td> 1 SpeI </td><td> 1 PstI </td><td> 2,5
| |
| - |
| |
| - | </td></tr></table>
| |
| - | <p>Digestions were incubated at 37°C for 3 hours, gel run and gel-extracted.
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| - | </p><p>Ligations were performed ON at 16°C:
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| - | </p>
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| - | <ul><li>I7: <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23118'>BBa_J23118</A> (S-P)+ I6 (X-P)
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| - | </li><li>I8: <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23110'>BBa_J23110</A> (S-P)+ I6 (X-P)
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| - | </li><li>I9: <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23114'>BBa_J23114</A> (S-P)+ I6 (X-P)
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| - | </li><li>I10: <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23116'>BBa_J23116</A> (S-P)+ I6 (X-P)
| |
| - |
| |
| - | </li></ul>
| |
| - | <p>NanoDrop quantifications were not reliable, so every ligation was performed with 3ul of vector, 2ul of insert, 3ul of ddH20, 1ul of T4 ligase and 1ul of T4 ligase buffer.
| |
| - | </p><p>These four promoters from Anderson Promoters Collection were chose for their <i>strength</i>, measured in Arbitrary Units:
| |
| - | </p>
| |
| - | <table border="1" align="center">
| |
| - | <tr>
| |
| - | <td> <b>Part</b> </td><td> <b>RFP (a.u.)</b>
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td><A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23118'>BBa_J23118</A></td><td>1429
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| - |
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td><A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23110'>BBa_J23110</A></td><td> 844
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| - | </td></tr>
| |
| - | <tr>
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| - | <td><A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23114'>BBa_J23114</A></td><td> 256
| |
| - | </td></tr>
| |
| - | <tr>
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| - | <td><A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_J23116'>BBa_J23116</A></td><td> 396
| |
| - | </td></tr>
| |
| - | </table>
| |
| | | | |
| - | <p>Soon quantitative tests will be performed to quantify each promoter's strength in terms of RPU.
| |
| - | </p><p>Today we received 2 stabs from iGEM HQ for <A HREF='http://partsregistry.org/wiki/index.php/Part:BBa_K208001'>BBa_K208001</A> (one for each registry location of this part), the Silver-fusion compatible BioBrick part that codes for phasin. This part is in a pSB3K3 plasmid. Bacteria were streaked on LB+Kan (50 ug/ml) agar plates. Cultures were named PhaP-1 and PhaP-2. Plates were incubated ON at 37°C.
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| - | </p>
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| | <div align="right"><small><a href="#indice" title="">^top</a></small></div> | | <div align="right"><small><a href="#indice" title="">^top</a></small></div> |
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| - | <td align="right"><a href="/Team:UNIPV-Pavia/Calendar/July/settimana1" title="Team:UNIPV-Pavia/Calendar/July/settimana1"> Next week</a></td> | + | <td align="right"><a href="/Team:UNIPV-Pavia/Calendar/July/settimana1" title="Team:UNIPV-Pavia/Calendar/June/settimana5"> Next week</a></td> |
| | | | |
| | </tr> | | </tr> |
"
