http://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&feed=atom&action=history2014hs.igem.org/team:Charlottesville RS/Notebook/041814 - Revision history2024-03-28T09:21:56ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&diff=22658&oldid=prevIngamename at 14:01, 20 June 20142014-06-20T14:01:02Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/material"><span><span>Material & Methods</span></span></a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/material"><span><span>Material & Methods</span></span></a></li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]--></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview" style="color: white">Notebook<!--[if gt IE 6]><!--></a><!--<![endif]--></div></td></tr>
</table>Ingamenamehttp://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&diff=8336&oldid=prevIngamename at 14:40, 2 June 20142014-06-02T14:40:47Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2014hs.igem.org/Team:Charlottesville_RS/Project"><span><span>Overall Project</span></span></a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2014hs.igem.org/Team:Charlottesville_RS/Project"><span><span>Overall Project</span></span></a></li></div></td></tr>
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</table>Ingamenamehttp://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&diff=5287&oldid=prevIngamename: /* April 18th, 2014 */2014-05-05T13:24:01Z<p><span class="autocomment">April 18th, 2014</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''Alli Ambrosini''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>''Alli Ambrosini''</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Did research on how <del class="diffchange diffchange-inline">we will be testing </del>the results of our project <del class="diffchange diffchange-inline">in class today</del>. <del class="diffchange diffchange-inline">We </del>will need to get non-nutrient agar plates so that <del class="diffchange diffchange-inline">we </del>can <del class="diffchange diffchange-inline">test </del>whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Did research on how <ins class="diffchange diffchange-inline">to test </ins>the results of our project. <ins class="diffchange diffchange-inline">The team </ins>will need to get non-nutrient agar plates so that <ins class="diffchange diffchange-inline">it </ins>can <ins class="diffchange diffchange-inline">be tested </ins>whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB. </div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">We took </del>competent cells, centrifuged them to get a pellet. <del class="diffchange diffchange-inline">We resuspended </del>in Luria broth, centrifuged and then <del class="diffchange diffchange-inline">we </del>combined into three tubes and added calcium chloride and put on ice for 20 minutes. <del class="diffchange diffchange-inline">Then </del>Alli <del class="diffchange diffchange-inline">took over after that step. She </del>centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Took </ins>competent cells, centrifuged them to get a pellet. <ins class="diffchange diffchange-inline">Resuspended </ins>in Luria broth, centrifuged and then combined into three tubes and added calcium chloride and put on ice for 20 minutes. Alli centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Did the re-suspending of DNA and transformation procedure.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Did the re-suspending of DNA and transformation procedure.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">1. With a pipette tip</del>, <del class="diffchange diffchange-inline">punch </del>a <del class="diffchange diffchange-inline">hole through the foil cover into the corresponding well of the Biobrick-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between </del>the <del class="diffchange diffchange-inline">wells.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The cells were possibly not competent because they were not suspended</ins>, <ins class="diffchange diffchange-inline">there was </ins>a <ins class="diffchange diffchange-inline">pellet at </ins>the <ins class="diffchange diffchange-inline">bottom</ins></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>2<del class="diffchange diffchange-inline">. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes </del>to <del class="diffchange diffchange-inline">make sure the dried DNA is fully re-suspended.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The transformation procedure in the iGEM procedure notebook on page 28 was performed There were not </ins>2 <ins class="diffchange diffchange-inline">hours </ins>to <ins class="diffchange diffchange-inline">incubate it, which was step 6 </ins></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">3</del>. <del class="diffchange diffchange-inline">Transform 5uL of </del>the <del class="diffchange diffchange-inline">resuspended </del>DNA <del class="diffchange diffchange-inline">into your competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Mrs</ins>. <ins class="diffchange diffchange-inline">Minutella plated </ins>the <ins class="diffchange diffchange-inline">transformed </ins>DNA at <ins class="diffchange diffchange-inline">60uL </ins>on <ins class="diffchange diffchange-inline">each plate</ins></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">4. We are going to use the chloramphenicol plates. Be sure to label them.</del></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: #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: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">We weren’t sure the cells were competent because they weren’t suspended, there was a pellet </del>at <del class="diffchange diffchange-inline">the bottom</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 class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">We then did the transformation procedure in the iGEM procedure notebook </del>on <del class="diffchange diffchange-inline">page 28. We did not have 2 hours to incubate it, which was step 6</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 class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]</div></td></tr>
</table>Ingamenamehttp://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&diff=5267&oldid=prevIngamename: /* April 18th, 2014 */2014-05-05T13:11:20Z<p><span class="autocomment">April 18th, 2014</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We then did the transformation procedure in the iGEM procedure notebook on page 28. We did not have 2 hours to incubate it, which was step 6</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We then did the transformation procedure in the iGEM procedure notebook on page 28. We did not have 2 hours to incubate it, which was step 6</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;"></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 style="color: red; font-weight: bold; text-decoration: none;">[[https://2014hs.igem.org/Team:Charlottesville_RS/Notebook/Overview Back]]</ins></div></td></tr>
</table>Ingamenamehttp://2014hs.igem.org/wiki/index.php?title=2014hs.igem.org/team:Charlottesville_RS/Notebook/041814&diff=4698&oldid=prevIngamename: /* April 18th, 2014 */2014-05-02T13:01:21Z<p><span class="autocomment">April 18th, 2014</span></p>
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<td colspan="2" class="diff-lineno">Line 423:</td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Did the re-suspending of DNA and transformation procedure.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Did the re-suspending of DNA and transformation procedure.</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;"></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.</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;"></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully re-suspended.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully re-suspended.</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;"></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3. Transform 5uL of the resuspended DNA into your competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3. Transform 5uL of the resuspended DNA into your competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.</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;"></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>4. We are going to use the chloramphenicol plates. Be sure to label them.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>4. We are going to use the chloramphenicol plates. Be sure to label them.</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>
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==April 18th, 2014==<br />
<br />
''Alli Ambrosini''<br />
<br />
Did research on how we will be testing the results of our project in class today. We will need to get non-nutrient agar plates so that we can test whether the bacteria can eat our PHB at all. Normal agar has dextrose in it, which bacteria will consume before they touch PHB because its easier to eat. A simple sugar is a lot easier to break down than a long polymer such as PHB. <br />
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''Lauren Ewell, John Grammer''<br />
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We took competent cells, centrifuged them to get a pellet. We resuspended in Luria broth, centrifuged and then we combined into three tubes and added calcium chloride and put on ice for 20 minutes. Then Alli took over after that step. She centrifuged gently by pulsing 10 times and then resuspended with 200uL of cold calcium chloride.<br />
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''Alex Manchester, Nick Keen, Anders Beaurline, Jessica Prax, Bailey Fernandez''<br />
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Did the re-suspending of DNA and transformation procedure.<br />
1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick-standard part that you want. Make sure you have properly oriented the plate. We recommend that you do not remove the foil cover, as it could lead to cross contamination between the wells.<br />
2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully re-suspended.<br />
3. Transform 5uL of the resuspended DNA into your competent cells, plate your transformation with the appropriate antibiotic* and grow overnight.<br />
4. We are going to use the chloramphenicol plates. Be sure to label them.<br />
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We weren’t sure the cells were competent because they weren’t suspended, there was a pellet at the bottom<br />
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We then did the transformation procedure in the iGEM procedure notebook on page 28. We did not have 2 hours to incubate it, which was step 6</div>Ingamename