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Team:Utah State/Notebook - Revision history
2024-03-28T13:01:20Z
Revision history for this page on the wiki
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http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=195746&oldid=prev
Cwbarentine at 03:24, 27 September 2013
2013-09-27T03:24:40Z
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Cwbarentine
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=195742&oldid=prev
Cwbarentine at 03:24, 27 September 2013
2013-09-27T03:24:20Z
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Cwbarentine
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=175545&oldid=prev
Skiracer96 at 23:00, 25 September 2013
2013-09-25T23:00:27Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 23:00, 25 September 2013</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Follow proceedure from step 7 in: </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Follow proceedure from step 7 in: </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>http://www.piercenet.com/instructions/2162203.pdf</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>http://www.piercenet.com/instructions/2162203.pdf</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><ins class="diffchange diffchange-inline"><br></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><ins class="diffchange diffchange-inline"> </p></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"> </div></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"><a name="Protein Gel"><div class="Header1"></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"> Protein Gel</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"> </div></a></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"> <div class="subHead1"></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"> <p class="textProfile3"></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">1. Heat treat protein fractions for 2-3min in boiling water with SDS and loading dye in 1:1 ratio (around 30 uL total volume).</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"><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">2.Load 10uL of Marker into gel and then load fractions.</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"><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">3. Run gel for 45-60 min depending on the migration of the dye front.</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"><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">4. Stain with coomassie overnight.</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"><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">5. De-stain gel the next day in DI water. </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"><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">6. Take photograph of gel.</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"><br> <br></ins></div></td></tr>
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=175491&oldid=prev
Skiracer96 at 22:54, 25 September 2013
2013-09-25T22:54:23Z
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div style="position: absolute; top: 150px; display: inline-block; float: left;"> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div style="position: absolute; top: 150px; display: inline-block; float: left;"> </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>5. Transfer supernatent to another tube.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>5. Transfer supernatent to another tube.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>6. Follow proceedure from step 7 in <del class="diffchange diffchange-inline"> <li> <a> href="</del>http://www.piercenet.com/instructions/2162203.pdf<del class="diffchange diffchange-inline">"Nickel column protocol </a> </li></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>6. Follow proceedure from step 7 in<ins class="diffchange diffchange-inline">: </ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>http://www.piercenet.com/instructions/2162203.pdf</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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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=175428&oldid=prev
Skiracer96 at 22:44, 25 September 2013
2013-09-25T22:44:19Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 22:44, 25 September 2013</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>5. Transfer supernatent to another tube.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>5. Transfer supernatent to another tube.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>6. Follow proceedure from step 7 in <li><del class="diffchange diffchange-inline">Nickel column protocol </del><a href="http://www.piercenet.com/instructions/2162203.pdf"<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>6. Follow proceedure from step 7 in <li> <a<ins class="diffchange diffchange-inline">> </ins>href="http://www.piercenet.com/instructions/2162203.pdf"<ins class="diffchange diffchange-inline">Nickel column protocol </ins> </a> </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>
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=175418&oldid=prev
Skiracer96 at 22:42, 25 September 2013
2013-09-25T22:42:10Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 22:42, 25 September 2013</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></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;"><a name="Protein purification"><div class="Header1"></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;"> Protein purification</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;"> </div></a></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;"> <div class="subHead1"></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;"> <p class="textProfile3"></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;"> Nickel columns, HisPur™ Ni-NTA Spin Columns (Catalog number: 88224), Thermofisher Scientific. Prior to running protein fractions through Nickel column the following modifications to the standard procedure was used:</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;">1.Add equilibrium buffer 3:1 ratio to mass of wet cell pellet, transfer to beaker.</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;"><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 style="color: red; font-weight: bold; text-decoration: none;">2. Sonicate for 5 min on max setting.</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;"><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 style="color: red; font-weight: bold; text-decoration: none;">3. Transfer 10 15mL tube and centrifuge for 10min @ 3500 rpm.</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;"><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 style="color: red; font-weight: bold; text-decoration: none;">4. Supernatent is your protein mix.</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;"><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 style="color: red; font-weight: bold; text-decoration: none;">5. Transfer supernatent to another tube.</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;"><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 style="color: red; font-weight: bold; text-decoration: none;">6. Follow proceedure from step 7 in <li>Nickel column protocol <a href="http://www.piercenet.com/instructions/2162203.pdf"> </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 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;"></ins></div></td></tr>
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=165980&oldid=prev
Skiracer96 at 05:18, 25 September 2013
2013-09-25T05:18:47Z
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=165979&oldid=prev
Skiracer96 at 05:18, 25 September 2013
2013-09-25T05:18:25Z
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=165975&oldid=prev
Skiracer96 at 05:18, 25 September 2013
2013-09-25T05:18:00Z
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Skiracer96
http://2013.igem.org/wiki/index.php?title=Team:Utah_State/Notebook&diff=149011&oldid=prev
Cwbarentine at 19:56, 22 September 2013
2013-09-22T19:56:06Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 19:56, 22 September 2013</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2013.igem.org/Team:Utah_State/Results"><img src="https://static.igem.org/mediawiki/2013/a/a9/ResultsText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/c/c6/ResultsSqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/a/a9/ResultsText.png'" width="233px" height="32px"></a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2013.igem.org/Team:Utah_State/Results"><img src="https://static.igem.org/mediawiki/2013/a/a9/ResultsText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/c/c6/ResultsSqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/a/a9/ResultsText.png'" width="233px" height="32px"></a></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2013.igem.org/Team:Utah_State/Notebook"><img src="https://static.igem.org/mediawiki/2013/3/32/ProtocolsText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/5/5e/ProtocolsSqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/3/32/ProtocolsText.png'" width="233px" height="32px"></a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2013.igem.org/Team:Utah_State/Notebook"><img src="https://static.igem.org/mediawiki/2013/3/32/ProtocolsText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/5/5e/ProtocolsSqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/3/32/ProtocolsText.png'" width="233px" height="32px"></a></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://2013.igem.org/Team:Utah_State/Notebook#Electroporation">Electroporation of <i>E. Coli</i></a></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <a href="https://2013.igem.org/Team:Utah_State/Notebook#Electroporation">Electroporation of <i>E. Coli</i></a></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 style="color: red; font-weight: bold; text-decoration: none;"> <a href="https://2013.igem.org/Team:Utah_State/Attributions"><img src="https://static.igem.org/mediawiki/2013/5/53/AttribText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/6/6e/AttribSqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/5/53/AttribText.png'" width="245px" height="32px"></a></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;"> <div style="margin-left: 5px;"><a href="https://2013.igem.org/Team:Utah_State/Safety"><img src="https://static.igem.org/mediawiki/2013/2/26/SafetyText.png" onMouseOver="this.src='https://static.igem.org/mediawiki/2013/a/ab/SafetySqgl.png'" onMouseOut= "this.src='https://static.igem.org/mediawiki/2013/2/26/SafetyText.png'" width="245px" height="32px"></a></div></ins></div></td></tr>
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Cwbarentine