http://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&feed=atom&action=historyTeam:Yale/Our Project/Notebook/Week 5 - Revision history2024-03-28T15:42:03ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=206111&oldid=prevLitagaki at 02:51, 28 October 20102010-10-28T02:51:19Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 7">week 7</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 7">week 7</a></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 8">week 8</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 8">week 8</a></li></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><li id="nb"><a href="https://2010.igem.org/Team:Yale/Our Project/Notebook/Week 9">week 9</a></li></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li><a href="https://2010.igem.org/Team:Yale/Our Project/Protocols">protocols</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li><a href="https://2010.igem.org/Team:Yale/Our Project/Protocols">protocols</a></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li><a href="https://2010.igem.org/Team:Yale/Our Project/Safety">safety</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li><a href="https://2010.igem.org/Team:Yale/Our Project/Safety">safety</a></li></div></td></tr>
</table>Litagakihttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=200795&oldid=prevDzhu at 23:34, 27 October 20102010-10-27T23:34:26Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Based on the results, all enzymes appeared to work, as they successfully created a linearized 3.2 kb fragment.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Based on the results, all enzymes appeared to work, as they successfully created a linearized 3.2 kb fragment.</li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 54-57 of the hard copy lab notebook. </i></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 54-57 of the hard copy lab notebook. </i></div></td></tr>
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</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=197772&oldid=prevDzhu at 21:45, 27 October 20102010-10-27T21:45:51Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Needed more phsABC, so ran five PCR reactions to amplify it from plasmid pSB74. Reaction mixture contents were the same as the DMSO variant used on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_4">6/30 </a>as was the thermocycler protocol (phs50). The results were then run on a gel</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Needed more phsABC, so ran five PCR reactions to amplify it from plasmid pSB74. Reaction mixture contents were the same as the DMSO variant used on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_4">6/30 </a>as was the thermocycler protocol (phs50). The results were then run on a gel</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Gel of PCR amplified phsABC Extreme right lane:1 kb ladder, all other lanes PCR reaction mixtures and their spillover</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><div id="caption"></ins>Gel of PCR amplified phsABC Extreme right lane:1 kb ladder, all other lanes PCR reaction mixtures and their spillover<ins class="diffchange diffchange-inline"></div></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Needed chloramphenicol plates for pSB1C3-based constructs, so mixed 350 μL of 50 μg/μL chloramphenicol with an autoclaved solution of 7.5 g agar and 12.5 g LB in 500 mL H2O and poured plates. </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Needed chloramphenicol plates for pSB1C3-based constructs, so mixed 350 μL of 50 μg/μL chloramphenicol with an autoclaved solution of 7.5 g agar and 12.5 g LB in 500 mL H2O and poured plates. </li></div></td></tr>
</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=197678&oldid=prevDzhu at 21:42, 27 October 20102010-10-27T21:42:10Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>The diagnostic 1.0% agarose gel was run at 90 V vs. a 1 kb ladder. Each of the thirteen samples gave one diffuse band around 3 kb and no evidence of the 3.6 kb phsABC fragment, so both ligation attempts #1 and #2 were unsuccessful.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>The diagnostic 1.0% agarose gel was run at 90 V vs. a 1 kb ladder. Each of the thirteen samples gave one diffuse band around 3 kb and no evidence of the 3.6 kb phsABC fragment, so both ligation attempts #1 and #2 were unsuccessful.</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Gel of XbaI and SpeI double digest of plasmids from ligation attempts #1 and #2 Lane 1: 1 kb ladder, Lanes 2-5:1A-4A, Lanes 6-14: 1B-9B <<del class="diffchange diffchange-inline">br</del>/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><div id="caption"></ins>Gel of XbaI and SpeI double digest of plasmids from ligation attempts #1 and #2 Lane 1: 1 kb ladder, Lanes 2-5:1A-4A, Lanes 6-14: 1B-9B </<ins class="diffchange diffchange-inline">div</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>During digestion prepped plasmid samples for sequencing. Each sample contained 3 μL of plasmid soln (average concentration of 96 ng/μL), 2 μL of 4 mM VF2 (forward primer), and 13 μL of distilled water. Intended to follow up with reverse sequencing of more promising samples, but gel results made it a moot point. </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>During digestion prepped plasmid samples for sequencing. Each sample contained 3 μL of plasmid soln (average concentration of 96 ng/μL), 2 μL of 4 mM VF2 (forward primer), and 13 μL of distilled water. Intended to follow up with reverse sequencing of more promising samples, but gel results made it a moot point. </li></div></td></tr>
</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=197648&oldid=prevDzhu at 21:40, 27 October 20102010-10-27T21:40:39Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <div id="caption>Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order</div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <div id="caption<ins class="diffchange diffchange-inline">"</ins>>Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order</div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol</a> </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol</a> </li></div></td></tr>
</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=197613&oldid=prevDzhu at 21:38, 27 October 20102010-10-27T21:38:51Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="caption>Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order</div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id="caption>Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order</div></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol<del class="diffchange diffchange-inline">.</del></a> </li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol</a> </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>This day's activities are also recorded on page 47 and 48 of the hard copy lab notebook. </i></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>This day's activities are also recorded on page 47 and 48 of the hard copy lab notebook. </i></div></td></tr>
</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=197579&oldid=prevDzhu at 21:37, 27 October 20102010-10-27T21:37:51Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ran colony PCR of colonies 1B-9B from ligation attempt #2 according to the same protocol as used on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_4"> 7/1</a>.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Ran colony PCR of colonies 1B-9B from ligation attempt #2 according to the same protocol as used on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_4"> 7/1</a>.</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Then ran a 1.0% agarose gel of the nine PCR reaction solutions against a 1 kb ladder, but the resulting gel failed to show any evidence of either insert or vector, so will have to rely on sequencing and digests of miniprepped plasmids.</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Then ran a 1.0% agarose gel of the nine PCR reaction solutions against a 1 kb ladder, but the resulting gel failed to show any evidence of either insert or vector, so will have to rely on sequencing and digests of miniprepped plasmids.</li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order<<del class="diffchange diffchange-inline">br</del>/></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><div align="center"></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <ins class="diffchange diffchange-inline"><div id="caption></ins>Gel of colony PCR from ligation attempt #2 Lane 1: 1 kb ladder, Lanes 3-11:PCR products from colonies 1B-9B, in order</<ins class="diffchange diffchange-inline">div</ins>></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol.</a> </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li>Miniprepped liquid cultures grown from colonies of first ligation attempt (colonies 1A-4A) and second ligation attempt (colonies 1B-9B) according to <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols"> standard miniprep protocol.</a> </li></div></td></tr>
</table>Dzhuhttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=188301&oldid=prevLitagaki at 16:48, 27 October 20102010-10-27T16:48:16Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Realize that mistakenly plated triple ligation reaction on ampicillin plate rather than the required chloramphenicol, so none of the colonies present are of value. Fortunately still have transformation solution, so replate it, this time on a chloramphenicol </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Realize that mistakenly plated triple ligation reaction on ampicillin plate rather than the required chloramphenicol, so none of the colonies present are of value. Fortunately still have transformation solution, so replate it, this time on a chloramphenicol </li></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><li> Inoculated overnight liquid cultures of ligation attempt #4 transformants (#9-#24 on index plates) for miniprep the following morning </li></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Growth assay prep work </b> <br/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Growth assay prep work </b> <br/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Want to see if pSB74 confers any copper resistance on <i>E. coli</i>, since it should help them precipitate nearby copper and thus reduce the copper concentration that they experience. </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Want to see if pSB74 confers any copper resistance on <i>E. coli</i>, since it should help them precipitate nearby copper and thus reduce the copper concentration that they experience. </li></div></td></tr>
</table>Litagakihttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=187624&oldid=prevLitagaki at 16:23, 27 October 20102010-10-27T16:23:32Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 16:23, 27 October 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Want to see if pSB74 confers any copper resistance on <i>E. coli</i>, since it should help them precipitate nearby copper and thus reduce the copper concentration that they experience. </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Want to see if pSB74 confers any copper resistance on <i>E. coli</i>, since it should help them precipitate nearby copper and thus reduce the copper concentration that they experience. </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Inoculated liquid cultures of LE392 with pSB74 in ampicillin LB and untransformed LE392 in plain LB and left to grow overnight on shaker at 37˚C for growth assays the following day </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li> Inoculated liquid cultures of LE392 with pSB74 in ampicillin LB and untransformed LE392 in plain LB and left to grow overnight on shaker at 37˚C for growth assays the following day </li></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><li> Prepared CuSO<sub>4</sub> solutions in LB. Each well in the 96-well plate will have 225 uL of copper solution and 25 uL of cell solution, so the copper solution must have CuSO<sub>4</sub><del class="diffchange diffchange-inline">, IPTG, and ampicillin concentrations </del>that <del class="diffchange diffchange-inline">are </del>10/9 the desired final values. <del class="diffchange diffchange-inline">Target </del>IPTG <del class="diffchange diffchange-inline">concentration is 2 mM, so start </del>at <del class="diffchange diffchange-inline">2.2 mM, and ampicillin concentration should </del>be <del class="diffchange diffchange-inline">100 ug/mL, so start at 111 ug/mL</del>. <del class="diffchange diffchange-inline"> </del>The desired CuSO<sub>4</sub> concentrations are those used in the wide and narrow concentration range trials on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook"> 6/10 </a> and <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_2"> 6/14 </a>. </li></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><li> Prepared CuSO<sub>4</sub> solutions in LB. Each well in the 96-well plate will have 225 uL of copper solution and 25 uL of cell solution, so the copper solution must have <ins class="diffchange diffchange-inline">a </ins>CuSO<sub>4</sub> <ins class="diffchange diffchange-inline">concentration </ins>that <ins class="diffchange diffchange-inline">is </ins>10/9 the desired final values. <ins class="diffchange diffchange-inline">The copper solutions must also contain ampicillin and </ins>IPTG at <ins class="diffchange diffchange-inline">the same levels as the cultures that will </ins>be <ins class="diffchange diffchange-inline">introduced</ins>. The desired CuSO<sub>4</sub> concentrations are those used in the wide and narrow concentration range trials on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook"> 6/10 </a> and <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_2"> 6/14 </a>. </li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 59-61 of the hard copy lab notebook.</i></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 59-61 of the hard copy lab notebook.</i></div></td></tr>
</table>Litagakihttp://2010.igem.org/wiki/index.php?title=Team:Yale/Our_Project/Notebook/Week_5&diff=187335&oldid=prevLitagaki at 16:13, 27 October 20102010-10-27T16:13:53Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 16:13, 27 October 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Sunday 7/11--</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Sunday 7/11--<ins class="diffchange diffchange-inline">Prep for copper growth assays of transformants</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a id="link" href="javascript:ReverseDisplay('sunday')">See more/less</a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><a id="link" href="javascript:ReverseDisplay('sunday')">See more/less</a></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="display:none;" id="sunday"></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="display:none;" id="sunday"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!------------- Sunday -------------></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!------------- Sunday -------------></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><<del class="diffchange diffchange-inline">h4</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><<ins class="diffchange diffchange-inline">b>Ligation work </b> <br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><ul></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li> Realize that mistakenly plated triple ligation reaction on ampicillin plate rather than the required chloramphenicol, so none of the colonies present are of value. Fortunately still have transformation solution, so replate it, this time on a chloramphenicol </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><b>Growth assay prep work </b> <br/></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li> Want to see if pSB74 confers any copper resistance on <i>E. coli</i>, since it should help them precipitate nearby copper and thus reduce the copper concentration that they experience. </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li> Inoculated liquid cultures of LE392 with pSB74 in ampicillin LB and untransformed LE392 in plain LB and left to grow overnight on shaker at 37˚C for growth assays the following day </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><li> Prepared CuSO<sub>4</sub> solutions in LB. Each well in the 96-well plate will have 225 uL of copper solution and 25 uL of cell solution, so the copper solution must have CuSO<sub>4</sub>, IPTG, and ampicillin concentrations that are 10/9 the desired final values. Target IPTG concentration is 2 mM, so start at 2.2 mM, and ampicillin concentration should be 100 ug/mL, so start at 111 ug/mL. The desired CuSO<sub>4</sub> concentrations are those used in the wide and narrow concentration range trials on <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook"> 6/10 </a> and <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_2"> 6/14 </a>. </li></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"></ul</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 59-61 of the hard copy lab notebook.</i></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><i>Wetlab work for this day is also recorded on pages 59-61 of the hard copy lab notebook.</i></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!------------- Sunday -------------></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><!------------- Sunday -------------></div></td></tr>
</table>Litagaki