http://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&feed=atom&action=historyTeam:Nevada/ccdB - Revision history2024-03-29T10:00:28ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=193497&oldid=prevHilarya: /* ccdB */2010-10-27T19:29:02Z<p><span class="autocomment">ccdB</span></p>
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<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: #ffa; color:black; font-size: smaller;"><div><html><a href="https://static.igem.org/mediawiki/2010/a/a0/Randy_and_Vadim.jpg"><img src="https://static.igem.org/mediawiki/2010/a/a0/Randy_and_Vadim.jpg" class="shadow" style="float:left;width:400px;margin:10px"></a></html><p>The ccdB gene is a known topoisomerase II poison and will kill most commercially available E. coli cell lines. The ccdB gene can be used as a selectable marker by placing it in the multicloning region of a plasmid and propagating it in ccdB resistant cell lines (e.g. DB3.1 from New England Biolabs). During cloning, the ccdB gene can be switched out for your gene of interest and transformed into a cell line that is susceptible to the toxic effects of ccdB (e.g NEBβ cells from New England Biolabs). Colonies that grow on plates should contain the plasmid and your gene of interest. If the ccdB gene is still present in the plasmid, the plasmid will kill any colonies where it is still present. This is an improvement to the current ccdB cell death gene part (BBa_P1010) as it does not contain the inactive ccdA gene or an unknown stuffer region.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><html><a href="https://static.igem.org/mediawiki/2010/a/a0/Randy_and_Vadim.jpg"><img src="https://static.igem.org/mediawiki/2010/a/a0/Randy_and_Vadim.jpg" class="shadow" style="float:left;width:400px;margin:10px"></a></html><p>The ccdB gene is a known topoisomerase II poison and will kill most commercially available E. coli cell lines. The ccdB gene can be used as a selectable marker by placing it in the multicloning region of a plasmid and propagating it in ccdB resistant cell lines (e.g. DB3.1 from New England Biolabs). During cloning, the ccdB gene can be switched out for your gene of interest and transformed into a cell line that is susceptible to the toxic effects of ccdB (e.g<ins class="diffchange diffchange-inline">. </ins>NEBβ cells from New England Biolabs). Colonies that grow on plates should contain the plasmid and your gene of interest. If the ccdB gene is still present in the plasmid, the plasmid will kill any colonies where it is still present. This is an improvement to the current ccdB cell death gene part (BBa_P1010) as it does not contain the inactive ccdA gene or an unknown stuffer region.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*ccdB was transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Left). </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*ccdB was transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Left). </div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*PCR gel confirming the presence of ccdB in amplified sample(Right). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*PCR gel confirming the presence of ccdB in amplified sample (Right). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
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</table>Hilaryahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=193311&oldid=prevEbersaba at 19:22, 27 October 20102010-10-27T19:22:36Z<p></p>
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</table>Ebersabahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=166016&oldid=prevRpares: /* ccdB */2010-10-26T20:34:13Z<p><span class="autocomment">ccdB</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>*ccdB was transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (<del class="diffchange diffchange-inline">Right</del>). </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*ccdB was transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (<ins class="diffchange diffchange-inline">Left</ins>). </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>*PCR gel confirming the presence of ccdB in amplified sample(<del class="diffchange diffchange-inline">Left</del>). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*PCR gel confirming the presence of ccdB in amplified sample(<ins class="diffchange diffchange-inline">Right</ins>). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
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</table>Rpareshttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=165995&oldid=prevRpares: /* ccdB */2010-10-26T20:32:55Z<p><span class="autocomment">ccdB</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</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>*ccdB <del class="diffchange diffchange-inline">in the PCR-Blunt II vector were </del>transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Right). </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>*ccdB <ins class="diffchange diffchange-inline">was </ins>transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Right). </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*PCR gel confirming the presence of ccdB in amplified sample(Left). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*PCR gel confirming the presence of ccdB in amplified sample(Left). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
</table>Rpareshttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=141290&oldid=prevMpolasko at 05:39, 25 October 20102010-10-25T05:39:29Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>'''Vectors Used:'''<html><a href="https://static.igem.org/mediawiki/2010/f/f4/Vectors_ccdB.png"><img src="https://static.igem.org/mediawiki/2010/f/f4/Vectors_ccdB.png" class="shadow" style="float:left;width:500px;margin:10px"></a></html<del class="diffchange diffchange-inline">></p</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>'''Vectors Used:'''<ins class="diffchange diffchange-inline"></p></ins><html><a href="https://static.igem.org/mediawiki/2010/f/f4/Vectors_ccdB.png"><img src="https://static.igem.org/mediawiki/2010/f/f4/Vectors_ccdB.png" class="shadow" style="float:left;width:500px;margin:10px"></a></html></div></td></tr>
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</table>Mpolaskohttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=139819&oldid=prevHilarya: /* ccdB */2010-10-25T01:35:03Z<p><span class="autocomment">ccdB</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><br>ccdB in the PCR-Blunt II vector were transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Right). </del></div></td><td colspan="2"> </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>PCR gel confirming the presence of ccdB in amplified sample(Left). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">*ccdB in the PCR-Blunt II vector were transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines (Right). </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">*</ins>PCR gel confirming the presence of ccdB in amplified sample(Left). <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
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</table>Hilaryahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=139813&oldid=prevHilarya: /* ccdB */2010-10-25T01:34:31Z<p><span class="autocomment">ccdB</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">(Right) </del>ccdB in the PCR-Blunt II vector were transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><br></ins>ccdB in the PCR-Blunt II vector were transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines <ins class="diffchange diffchange-inline">(Right)</ins>. </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><del class="diffchange diffchange-inline">(Left) </del>PCR gel confirming the presence of ccdB in amplified sample.<html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>PCR gel confirming the presence of ccdB in amplified sample<ins class="diffchange diffchange-inline">(Left)</ins>. <html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></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>
</table>Hilaryahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=139799&oldid=prevHilarya: /* ccdB */2010-10-25T01:33:30Z<p><span class="autocomment">ccdB</span></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 01:33, 25 October 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Method:''' ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.<html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>'''Our Method:''' ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.<html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><p>&nbsp;</p></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </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;"></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><p>'''Our Results:'''<html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html><del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>'''Our Results:'''</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>&nbsp;</p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">(Right) ccdB in the PCR-Blunt II vector were transformed in three different cell lines: NEB10β cells (New England Biolabs), Omni Max 2 cells (Invitrogen), and DB3.1 cells (New England Biolabs). NEB10β and Omni Max 2 cell lines are not resistant to the toxic properties of ccdB. No colonies were present in those two cell lines but were present in DB3.1 cell lines.</ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>&nbsp;</p></del></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><del class="diffchange diffchange-inline"><p>&nbsp;</p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">(Left) PCR gel confirming the presence of ccdB in amplified sample.</ins><html><a href="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png"><img src="https://static.igem.org/mediawiki/2010/6/63/Ccdb_results.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>&nbsp;</p></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><del class="diffchange diffchange-inline"><p>&nbsp;</del></p></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><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></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>----</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;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
</table>Hilaryahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=139793&oldid=prevHilarya: /* ccdB */2010-10-25T01:31:55Z<p><span class="autocomment">ccdB</span></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 01:31, 25 October 2010</td>
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<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;"></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><p>'''Our Method:'''ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.<html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html><del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>'''Our Method:''' ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.<html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></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"><p>&nbsp;</p></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><del class="diffchange diffchange-inline"><p>&nbsp;</p></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><del class="diffchange diffchange-inline"><p>&nbsp;</del></p></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><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>&nbsp;</p></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>----</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;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
</table>Hilaryahttp://2010.igem.org/wiki/index.php?title=Team:Nevada/ccdB&diff=139789&oldid=prevHilarya: /* ccdB */2010-10-25T01:31:34Z<p><span class="autocomment">ccdB</span></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 01:31, 25 October 2010</td>
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<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;"></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><p>'''Our Method:'''<html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html><del class="diffchange diffchange-inline"></p></del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>'''Our Method:'''<ins class="diffchange diffchange-inline">ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.</ins><html><a href="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png"><img src="https://static.igem.org/mediawiki/2010/0/05/CcdB_method.png" class="shadow" style="float:center;width:850px;margin:10px"></a></html></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"><p>ccdB was amplified using Polymerase Chain Reaction (PCR) with primers designed to contain the specific BioBrick prefix and suffix. The amplified fragment was then TOPO-cloned into a PCR-Blunt II vector (Invitrogen). The vector was then cut with EcoRI and PstI restriction enzymes and the ccdB fragment was transformed into the iGEM compatible vector pSB1C3.</p></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><del class="diffchange diffchange-inline"><p>&nbsp;</del></p></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;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></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>&nbsp;</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 style="color: red; font-weight: bold; text-decoration: none;"><p>&nbsp;</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 style="color: red; font-weight: bold; text-decoration: none;"><p>&nbsp;</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 style="color: red; font-weight: bold; text-decoration: none;"><p>&nbsp;</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 style="color: red; font-weight: bold; text-decoration: none;"><p>&nbsp;</p></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>
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</table>Hilarya