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Team:Caltech/week11
From 2014.igem.org
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<b><font size=+1>Week 11</font></b><br><br> | <b><font size=+1>Week 11</font></b><br><br> | ||
<b><a href='week12'>Week 12</a></b><br><br> | <b><a href='week12'>Week 12</a></b><br><br> | ||
| + | <b><a href='week13'>Week 13</a></b><br><br> | ||
| + | <b><a href='week14'>Week 14</a></b><br><br> | ||
| + | <b><a href='week15'>Week 15</a></b><br><br> | ||
<!-- links end here --> | <!-- links end here --> | ||
</td> | </td> | ||
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</td> | </td> | ||
<td valign="top"> | <td valign="top"> | ||
| + | <b>lamBCDA & fsrABC Reception Systems</b> | ||
| + | <ul> | ||
| + | </ul> | ||
| + | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
<ul> | <ul> | ||
</ul> | </ul> | ||
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</td> | </td> | ||
<td valign="top"> | <td valign="top"> | ||
| + | <b>lamBCDA & fsrABC Reception Systems</b> | ||
| + | <ul> | ||
| + | </ul> | ||
| + | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
<ul> | <ul> | ||
</ul> | </ul> | ||
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</td> | </td> | ||
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| + | <b>lamBCDA & fsrABC Reception Systems</b> | ||
| + | <ul> | ||
| + | </ul> | ||
| + | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
<ul> | <ul> | ||
</ul> | </ul> | ||
| - | |||
</td> | </td> | ||
</tr> | </tr> | ||
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| + | <b>lamBCDA & fsrABC Reception Systems</b> | ||
<ul> | <ul> | ||
| + | </ul> | ||
| + | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
| + | <ul><li>Re-ran a Gibson assembly of pAA009 </li> | ||
| + | <li>Transformed Gibson products into JM109 cells </li> | ||
</ul> | </ul> | ||
</td> | </td> | ||
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</td> | </td> | ||
<td valign="top"> | <td valign="top"> | ||
| + | <b>lamBCDA & fsrABC Reception Systems</b> | ||
<ul> | <ul> | ||
| + | </ul> | ||
| + | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
| + | <ul><li>Picked 10 Gibson colonies from yesterday </li> | ||
| + | <li>Ran colony PCR on re-suspended colonies, inoculated liquid cultures of picked colonies </li> | ||
| + | <li>Ran gel of colony PCR products, but no good bands were obtained </li> | ||
| + | </ul> | ||
| + | </td> | ||
| + | </tr> | ||
| + | |||
| + | <tr><td bgColor="#777777" colspan="2" height="1px"></td></tr> | ||
| + | |||
| + | <tr> | ||
| + | <td width="25%" valign="top"> | ||
| + | <h4>Saturday, 8/30/14</h4> | ||
| + | </td> | ||
| + | <td valign="top"> | ||
| + | <b>Export Systems</b> | ||
| + | <br>Redo of cloning of agr system: | ||
| + | <ul><li>Ran PCR reactions to construct appropriate backbone for pTG002 & modified gblock insert for pTG002.</li> | ||
| + | <li>DpnI digested PCR product for pTG002 backbone.</li> | ||
| + | <li>PCR purified DpnI digested pTG002 backbone and pTG002 modified gblock insert PCR products.</li> | ||
| + | <li>Ran purified products on a gel.</li> | ||
| + | <li>Ran Gibson assembly reaction between pTG002-backbone & pTG002-insert.</li> | ||
| + | <li>Transformed Gibson reaction mix into JM109. Cells were then plated & incubated overnight.</li> | ||
| + | </ul> | ||
| + | De-FLAGging pTG005 for LC/MS analysis: | ||
| + | <ul><li>Ran PCR reaction to exclude 3xFLAG from pTG005.</li> | ||
| + | <li>Ran DpnI digest and then PCR purification on PCR product.</li> | ||
| + | <li>Ran PCR product on a gel to verify creation of the appropriate DNA fragment.</li> | ||
| + | <li>Ran a round-the-horn ligation reaction on the purified PCR product.</li> | ||
| + | <li>Transformed the reaction mix into JM109. Cells were then plated & incubated overnight.</li> | ||
</ul> | </ul> | ||
</td> | </td> | ||
Latest revision as of 00:53, 15 September 2014
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