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JUNE: WEEK 5
June, 28thFirst time in wet lab 11 LB agar plates, 35 LB agar + Amp plates and 15 LB agar + Cm 12.5 were prepared.
For each of them 1 ul was transformed in 100 ul of home-made TOP10 competent cells. Transformants were plated on LB agar plates with the right antibiotic and incubated ON at 37°C. June, 29thIn the morning all plates showed colonies (<partinfo>BBa_J04450</partinfo> showed light red colonies); for each one we picked one colony and inoculated it in 1 ml of liquid LB with the proper antibiotic (ampicillin for <partinfo>BBa_C0060</partinfo>, <partinfo>BBa_C0061</partinfo> and <partinfo>BBa_K081022</partinfo>, kanamycin for <partinfo>BBa_J04450</partinfo>). Cultures were grown at 37°C, 220 rpm. In the afternoon glycerol stocks were prepared (750 ul of bacteria and 250 ul of 80% glycerol) and stored at -80°C except for <partinfo>BBa_C0061</partinfo> which was not sufficiently grown (we let it grow until next morning); then cultures were refilled with 5 ml of liquid LB with the proper antibiotic and incubated ON at 37°C, 220 rpm. June, 30thAll cultures were saturated; glycerol stock for <partinfo>BBa_C0061</partinfo> was prepared and stored at -80°C. Plasmids were extracted with MiniPrep kit; purified DNA was quantified with NanoDrop Spectrophotometer:
<partinfo>BBa_B0030</partinfo>, <partinfo>BBa_B0031</partinfo>, <partinfo>BBa_B0032</partinfo>, <partinfo>BBa_B0034</partinfo>, <partinfo>BBa_B0015</partinfo>, <partinfo>pSB4C5</partinfo>, <partinfo>BBa_I13501</partinfo>, <partinfo>BBa_I13507</partinfo> were inoculated from freezer glycerol stocks and grown ON at 37°C, 220 rpm for MiniPrep.
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Team:UNIPV-Pavia/Calendar/June/settimana5
From 2011.igem.org
(Difference between revisions)
| Line 28: | Line 28: | ||
<p> | <p> | ||
<ul type="circle"> | <ul type="circle"> | ||
| - | <li>< | + | <li></html><partinfo>BBa_C0060</partinfo><html> was resuspended from iGEM 2011 kit distribution, Plate 1, well 4K in 15 ul of ddH2O |
| - | <li>< | + | <li></html><partinfo>BBa_C0061</partinfo><html> was resuspended from iGEM 2011 kit distribution, Plate 1, well 4M in 15 ul of ddH2O |
| - | <li>< | + | <li></html><partinfo>BBa_J04450</partinfo><html> was resuspended from iGEM 2011 kit distribution, Plate 1, well 5E in 15 ul of ddH2O |
| - | <li>< | + | <li></html><partinfo>BBa_K081022</partinfo><html> was resuspended from iGEM 2011 kit distribution, Plate 2, well 12N in 15 ul of ddH2O |
</ul> | </ul> | ||
</p> | </p> | ||
| Line 44: | Line 44: | ||
<p> | <p> | ||
| - | In the morning all plates showed colonies (< | + | In the morning all plates showed colonies (</html><partinfo>BBa_J04450</partinfo><html> showed light red colonies); for each one we picked one colony and inoculated it in 1 ml of liquid LB with the proper antibiotic (ampicillin for </html><partinfo>BBa_C0060</partinfo><html>, </html><partinfo>BBa_C0061</partinfo><html> and </html><partinfo>BBa_K081022</partinfo><html>, kanamycin for </html><partinfo>BBa_J04450</partinfo><html>). Cultures were grown at 37°C, 220 rpm. |
</p> | </p> | ||
<p> | <p> | ||
| - | In the afternoon glycerol stocks were prepared (750 ul of bacteria and 250 ul of 80% glycerol) and stored at -80°C except for < | + | In the afternoon glycerol stocks were prepared (750 ul of bacteria and 250 ul of 80% glycerol) and stored at -80°C except for </html><partinfo>BBa_C0061</partinfo><html> which was not sufficiently grown (we let it grow until next morning); then cultures were refilled with 5 ml of liquid LB with the proper antibiotic and incubated ON at 37°C, 220 rpm. |
</p> | </p> | ||
<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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<a name="June.2C_30th"></a><h2> <span class="mw-headline">June, 30th</span></h2> | <a name="June.2C_30th"></a><h2> <span class="mw-headline">June, 30th</span></h2> | ||
<p> | <p> | ||
| - | All cultures were saturated; glycerol stock for < | + | All cultures were saturated; glycerol stock for </html><partinfo>BBa_C0061</partinfo><html> was prepared and stored at -80°C. |
</p> | </p> | ||
<p> | <p> | ||
| Line 68: | Line 68: | ||
<tr> | <tr> | ||
| - | <td>< | + | <td></html><partinfo>BBa_C0060</partinfo><html></td> |
<td>60.8</td> | <td>60.8</td> | ||
</tr> | </tr> | ||
| Line 74: | Line 74: | ||
<tr> | <tr> | ||
| - | <td>< | + | <td></html><partinfo>BBa_C0061</partinfo><html></td> |
<td>75.7</td> | <td>75.7</td> | ||
</tr> | </tr> | ||
| Line 80: | Line 80: | ||
<tr> | <tr> | ||
| - | <td>< | + | <td></html><partinfo>BBa_K081022</partinfo><html></td> |
<td>63.6</td> | <td>63.6</td> | ||
</tr> | </tr> | ||
| Line 86: | Line 86: | ||
<tr> | <tr> | ||
| - | <td>< | + | <td></html><partinfo>BBa_J04450</partinfo><html></td> |
<td>17.5</td> | <td>17.5</td> | ||
</tr> | </tr> | ||
| Line 93: | Line 93: | ||
<p> | <p> | ||
| - | < | + | </html><partinfo>BBa_B0030</partinfo><html>, </html><partinfo>BBa_B0031</partinfo><html>, </html><partinfo>BBa_B0032</partinfo><html>, </html><partinfo>BBa_B0034</partinfo><html>, </html><partinfo>BBa_B0015</partinfo><html>, </html><partinfo>pSB4C5</partinfo><html>, </html><partinfo>BBa_I13501</partinfo><html>, </html><partinfo>BBa_I13507</partinfo><html> were inoculated from freezer glycerol stocks and grown ON at 37°C, 220 rpm for MiniPrep. |
</p> | </p> | ||
<div align="right"><small><a href="#indice" title="">^top</a></small></div> | <div align="right"><small><a href="#indice" title="">^top</a></small></div> | ||
Revision as of 14:34, 16 July 2011
