http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&feed=atom&action=historyTeam:TU Delft/Project/rbs-characterization/parts - Revision history2024-03-28T11:04:35ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=194713&oldid=prevJcnossen: /* Cloning method */2010-10-27T20:09:25Z<p><span class="autocomment">Cloning method</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><i> </del>This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see what was done under the header 'Characterization of RBS sequences' in our <del class="diffchange diffchange-inline">blogs of June and July </del>2010.<del class="diffchange diffchange-inline"><</del>/<del class="diffchange diffchange-inline">i></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see what was done under the header 'Characterization of RBS sequences' in our <ins class="diffchange diffchange-inline">blog posts starting around</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">[https://</ins>2010.<ins class="diffchange diffchange-inline">igem.org</ins>/<ins class="diffchange diffchange-inline">Team:TU_Delft#page=Notebook/blog&blog=6_July_2010 6 July]</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td></tr>
</table>Jcnossenhttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=160082&oldid=prevPtmvanboheemen at 13:57, 26 October 20102010-10-26T13:57:27Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>I13401 SpecifiedComponents</partinfo></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>I13401 SpecifiedComponents</partinfo></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;"><html><center><img src="https://static.igem.org/mediawiki/2010/0/00/TU_Delft_project_navigation.jpg" usemap="#projectnavigation" border="0" /></center><map id="projectnavigation" name="projectnavigation"><area shape="rect" alt="Characterization" title="" coords="309,3,591,45" href="https://2010.igem.org/Team:TU_Delft#page=Project/rbs-characterization/characterization" target="" /><area shape="rect" alt="Results" title="" coords="609,3,891,44" href="https://2010.igem.org/Team:TU_Delft#page=Project/rbs-characterization/results" target="" /><area shape="rect" alt="Parts" title="" coords="9,3,290,44" href="https://2010.igem.org/Team:TU_Delft#page=Project/rbs-characterization/parts" target="" /></map></html></ins></div></td></tr>
</table>Ptmvanboheemenhttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130936&oldid=prevMvoges: /* Cloning method */2010-10-24T11:07:39Z<p><span class="autocomment">Cloning method</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><i> This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see <del class="diffchange diffchange-inline">the </del>what was done under the header 'Characterization of RBS sequences' in our blogs of June and July 2010.</i></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><i> This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see what was done under the header 'Characterization of RBS sequences' in our blogs of June and July 2010.</i></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130931&oldid=prevMvoges: /* Cloning method */2010-10-24T11:07:06Z<p><span class="autocomment">Cloning method</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><i> This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see the what was done under the header 'Characterization of RBS sequences' in our blogs of June and July 2010.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><i> This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see the what was done under the header 'Characterization of RBS sequences' in our blogs of June and July 2010.<ins class="diffchange diffchange-inline"></i></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130927&oldid=prevMvoges: /* Cloning method */2010-10-24T11:06:38Z<p><span class="autocomment">Cloning method</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Cloning method===</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;"><i> This is a summary of the cloning methods that succeeded. For more information on the exact methods and all attempts made see the what was done under the header 'Characterization of RBS sequences' in our blogs of June and July 2010.</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130871&oldid=prevMvoges: /* The Reference Construct */2010-10-24T10:55:58Z<p><span class="autocomment">The Reference Construct</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Reference Construct====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Reference Construct====</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_J23100 BBa_J23100], which is contained in the pSB1A2 plasmid was digested using SpeI and PstI. [http://partsregistry.org/Part:BBa_E0240 BBa_E0240] was digested using XbaI and PstI and ligated with the digest product of J23100. Transformation into <i>E.coli</i> TOP10 yielded positives as <del class="diffchange diffchange-inline">analyzed </del>by UV-light. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_J23100 BBa_J23100], which is contained in the pSB1A2 plasmid was digested using SpeI and PstI. [http://partsregistry.org/Part:BBa_E0240 BBa_E0240] was digested using XbaI and PstI and ligated with the digest product of J23100. Transformation into <i>E.coli</i> TOP10 yielded positives as <ins class="diffchange diffchange-inline">determined </ins>by UV-light.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The BioBricks====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The BioBricks====</div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130835&oldid=prevMvoges: /* Parts */2010-10-24T10:46:28Z<p><span class="autocomment">Parts</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== Parts ==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>== Parts ==</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">Our </del>constructs used for measurements:</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">The </ins>constructs used for measurements:</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>K398500 SpecifiedComponents</partinfo></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>K398500 SpecifiedComponents</partinfo></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>K173000 SpecifiedComponents</partinfo></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><partinfo>K173000 SpecifiedComponents</partinfo></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;"><partinfo>I13401 SpecifiedComponents</partinfo></ins></div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130832&oldid=prevMvoges: /* Parts */2010-10-24T10:45:41Z<p><span class="autocomment">Parts</span></p>
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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;"></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;"><partinfo>K173000 SpecifiedComponents</partinfo></ins></div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130829&oldid=prevMvoges: /* The BioBricks */2010-10-24T10:45:04Z<p><span class="autocomment">The BioBricks</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The BioBricks====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The BioBricks====</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The Biobricks generated for the measurements were: [http://partsregistry.org/Part:BBa_K398500 K398500], [http://partsregistry.org/Part:BBa_K398501 K398501], [http://partsregistry.org/Part:BBa_K398502 K398502], [http://partsregistry.org/Part:BBa_K398503 K398503], [http://partsregistry.org/Part:BBa_K398504 K398504] and [http://partsregistry.org/Part:BBa_K173000 K173000]. A general example map of the construction is shown below, where the RBS is displayed in fucsia.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The Biobricks generated for the measurements were: [http://partsregistry.org/Part:BBa_K398500 K398500], [http://partsregistry.org/Part:BBa_K398501 K398501], [http://partsregistry.org/Part:BBa_K398502 K398502], [http://partsregistry.org/Part:BBa_K398503 K398503], [http://partsregistry.org/Part:BBa_K398504 K398504] and [http://partsregistry.org/Part:BBa_K173000 K173000]. A general example map of the construction is shown below, where the RBS is displayed in fucsia<ins class="diffchange diffchange-inline">. Notice the additional XbaI site located between the promoter and RBS sequence. This site is inherent to the Anderson RBS inserts on the distribution plates and was maintained in our constructs in case promoter swapping would be necessary in future</ins>.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:RBS1.jpg|left|650px]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:RBS1.jpg|left|650px]]</div></td></tr>
</table>Mvogeshttp://2010.igem.org/wiki/index.php?title=Team:TU_Delft/Project/rbs-characterization/parts&diff=130800&oldid=prevMvoges: /* The Anderson Constructs */2010-10-24T10:42:01Z<p><span class="autocomment">The Anderson Constructs</span></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 10:42, 24 October 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Anderson Constructs====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[http://partsregistry.org/Part:BBa_I13401 BBa_I13401] was PCR amplified by using the universal primers G00100 and G00101. The purified PCR product was back-digested using XbaI and PstI. </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The Anderson RBSs are provided in pSB1A2 plasmids. These were digested using SpeI and PstI and ligated with the digested I13401 insert. Transformation into competent <del class="diffchange diffchange-inline">Top10 </del><i>E.coli</i> cells yielded positives, as determined by UV-light plate analysis.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The Anderson RBSs are provided in pSB1A2 plasmids. These were digested using SpeI and PstI and ligated with the digested I13401 insert. Transformation into competent <ins class="diffchange diffchange-inline">TOP10 </ins><i>E.coli</i> cells yielded positives, as determined by UV-light plate analysis.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Reference Construct====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====The Reference Construct====</div></td></tr>
</table>Mvoges