Team:Caltech/week5

From 2014.igem.org

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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 valign="top">
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<ul>
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<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 &mu;L of each of the 5 Gibson products was then transformed into JM109. The bacteria were then plated on 5 carbenicillin plates & incubated @ 37&deg;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
Home Team Official Team Profile Project Parts Modeling Notebook Safety Attributions
Notebook
Overview

Week 1

Week 2

Week 3

Week 4

Week 5

Week 6

Week 7

Week 8

Week 9

Week 10

Week 11

Week 12

Week Five

Monday, 7/14/14

 Export Systems
  • Redid Gibson assembly of comX constructs to reattempt cloning
  • Constructs transformed into JM109, plated on carbenicillin, and incubated overnight
lamBCDA & fsrABC Reception Systems
  • Grew liquid cultures of E. coli transformed with pWW2149+pWW1521 and pWW2149+pWW1523 (colonies had been allowed to grow on agar plates over the weekend)
agrBCDA Reception System and Combinatorial Promoters

Tuesday, 7/15/14

 Export Systems
  • ComX System
    • Ran colony PCR again on overnight colonies with reassembled comX constructs (3 colonies picked from each of the 5 plates)
    • Ran a gel on the PCR products. Resulting gel.
  • agrBD, lamBD, & FsrB Systems
    • PCR assembly of backbones with overhangs needed for assembly of agrBD, lamBD, & FsrB export systems
    • DpnI digestion of PCR product for 2 hours and then PCR purification were run to remove any non-backbone fragments of DNA
    • The five export constructs to test [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
    • 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
lamBCDA & fsrABC Reception Systems

Wednesday, 7/16/14

Thursday, 7/17/14

Friday, 7/18/14

Retrieved from "http://2014.igem.org/Team:Caltech/week5"