Team:Caltech/week5
From 2014.igem.org
(Difference between revisions)
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<ul><li>Redid Gibson assembly of comX constructs to reattempt cloning</li> | <ul><li>Redid Gibson assembly of comX constructs to reattempt cloning</li> | ||
<li>Constructs transformed into JM109, plated on carbenicillin, and incubated overnight</li> | <li>Constructs transformed into JM109, plated on carbenicillin, and incubated overnight</li> | ||
+ | </ul> | ||
+ | <b>lamBCDA & fsrABC Reception Systems</b> | ||
+ | <ul><li>Grew liquid cultures of <i>E. coli</i> transformed with pWW2149+pWW1521 and pWW2149+pWW1523 (colonies had been allowed to grow on agar plates over the weekend)</li> | ||
+ | </ul> | ||
+ | <b>agrBCDA Reception System and Combinatorial Promoters</b> | ||
+ | <ul><li></li> | ||
</ul> | </ul> | ||
</td> | </td> | ||
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</td> | </td> | ||
<td valign="top"> | <td valign="top"> | ||
- | <ul> | + | <b>Export Systems</b> |
+ | <ul><li>ComX System | ||
+ | <ul><li>Ran colony PCR again on overnight colonies with reassembled comX constructs (3 colonies picked from each of the 5 plates)</li> | ||
+ | <li>Ran a gel on the PCR products. <a href="2014.igem.org/Team:Caltech/Results">Resulting gel.</a></li> | ||
+ | </ul></li> | ||
+ | <li>agrBD, lamBD, & FsrB Systems | ||
+ | <ul><li>PCR assembly of backbones with overhangs needed for assembly of agrBD, lamBD, & FsrB export systems</li> | ||
+ | <li>DpnI digestion of PCR product for 2 hours and then PCR purification were run to remove any non-backbone fragments of DNA</li> | ||
+ | <li>The <a href="2014.igem.org/Team:Caltech/">five export constructs to test</a> [link to images of the 5 plasmids we constructed] were assembled via Gibson using the PCRed backbones and geneblocks (containing the inserts) that arrived yesterday afternoon</li> | ||
+ | <li>1 μL of each of the 5 Gibson products was then transformed into JM109. The bacteria were then plated on 5 carbenicillin plates & incubated @ 37°C overnight</li> | ||
+ | </ul></li> | ||
+ | </ul> | ||
+ | <b>lamBCDA & fsrABC Reception Systems</b> | ||
+ | <ul><li> | ||
</ul> | </ul> | ||
</td> | </td> |
Revision as of 17:37, 16 July 2014
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