Team:Yale/Notebook/Week11
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<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week8">Week 8</a></li> | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week8">Week 8</a></li> | ||
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week9">Week 9</a></li> | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week9">Week 9</a></li> | ||
- | <li><a | + | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week10">Week 10</a></li> |
- | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week11">Week 11</a></li> | + | <li><a id="page" href="https://2011.igem.org/Team:Yale/Notebook/Week11">Week 11</a></li> |
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week12">Week 12</a></li> | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week12">Week 12</a></li> | ||
<li><a href="https://2011.igem.org/Team:Yale/Notebook/Week13">Week 13</a></li> | <li><a href="https://2011.igem.org/Team:Yale/Notebook/Week13">Week 13</a></li> | ||
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<div id="entry"> | <div id="entry"> | ||
- | + | <h1>Week 11: July 24-31, 2011</h1> | |
- | + | <ul> | |
- | - Planned/mapped out next 4 weeks | + | <li>Monday:<ul> |
- | + | <li>Regenerate Nickel columns</li> | |
- | + | <li>Incubate sample with beads</li> | |
- | + | <li>Survival assay</li> | |
- | + | </ul> | |
- | + | Tuesday:<ul> | |
- | + | <li>Survival Assay again</li> | |
- | + | <li>Elution from loose beads + SDS</li> | |
- | + | </ul></li><li> | |
- | + | Wednesday:<ul> | |
- | + | <li>Run flow-through through beads again</li> | |
- | + | </ul></li><li> | |
- | + | Thursday:<ul> | |
- | + | <li>Researched ice-affinity purification</li> | |
- | + | <li>Dialyzed sample</li> | |
- | + | </ul></li><li> | |
- | + | Friday:<ul> | |
- | + | <li>Planned/mapped out next 4 weeks</li> | |
- | + | <li>Took dialyzed bag with purified eGFP-RiAFP and divided it into 12 eppendorfs, containing 1 mL each. Flash-froze tubes in liquid nitrogen, then quickly used a hot edge to poke holes into top of eppendorfs to allow sublimation. Tubes then placed in lyophilizer for 3 nights.</li> | |
- | + | </ul></li><li> | |
- | + | Saturday:<ul> | |
- | + | <li>Began cloning all pSB1AK8 constructs into pSB1C3 for parts submission (G3, H1, RiA, RiG, T2, Z2?) by digesting those 6 constructs and linearized pSB1C3 with EcoRI, PstI | |
- | + | </li> | |
- | + | <li>Also, PCR amplified eGFP segment from original eGFP-RiAFP plasmid to use as a control for survival assays</li> | |
- | + | <li>Digested PCR fragment with eGFP with XbaI, PstI, and digested BBa_I712074 (strong T7 promoter) with SpeI and PstI</li> | |
- | + | <li>Gel extracted all digested pSB1AK8 constructs using kit, gel visualized with SYBRSafe (digestion seemed successful)</li> | |
- | + | </ul></li><li> | |
- | + | Sunday:<ul> | |
- | + | <li>Ligated gel-extracted pSB1AK8 fragments with pSB1C3; ligated eGFP PCR fragment with BBa_I712074</li> | |
- | + | <li>Transformed ligations into DH5alpha cells, plated them</li> | |
- | + | </ul></li> | |
- | + | </ul> | |
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</div> | </div> | ||
<div style="clear:both"></div> | <div style="clear:both"></div> | ||
</div> | </div> |
Latest revision as of 05:24, 28 September 2011