http://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&feed=atom&action=historyTeam:HokkaidoU Japan/Notebook/aggregation Week 1 - Revision history2024-03-28T16:56:59ZRevision history for this page on the wikiMediaWiki 1.16.0http://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=230950&oldid=prevTaKeZo: /* Digestion of I719005, B0034 and pSB1K3 */2012-09-27T01:44:13Z<p><span class="autocomment">Digestion of I719005, B0034 and pSB1K3</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|13 ul</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|13 ul</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ethanol precipitation of Digestion products====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Ethanol precipitation of Digestion products====</div></td></tr>
</table>TaKeZohttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=230879&oldid=prevTaKeZo at 01:43, 27 September 20122012-09-27T01:43:05Z<p></p>
<a href="http://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=230879&oldid=214021">Show changes</a>TaKeZohttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=214021&oldid=prevSlecat at 20:11, 26 September 20122012-09-26T20:11:27Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transformation===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transformation===</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div>Transformation of BBa_B0015(dT), B0034(RBS), I179005(pT7), K346007(Ag43), and K542009(pLacI-RBS-Ag43) <del class="diffchange diffchange-inline">to </del>DH5&alpha;</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Transformation of BBa_B0015(dT), B0034(RBS), I179005(pT7), K346007(Ag43), and K542009(pLacI-RBS-Ag43) <ins class="diffchange diffchange-inline">into </ins>DH5&alpha;</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Plated them on LBA(dt,RBS,T7) and LBC(Ag43, pLacI-RBS-Ag43). ]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Plated them on LBA(dt,RBS,T7) and LBC(Ag43, pLacI-RBS-Ag43). ]</div></td></tr>
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<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>Transformation of K346007(Ag43) <del class="diffchange diffchange-inline">to </del>DH5&alpha;.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Transformation of K346007(Ag43) <ins class="diffchange diffchange-inline">into </ins>DH5&alpha;.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#To gain chrolamphenicol resistance solution was pre-incubated for 2 hrs.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#To gain chrolamphenicol resistance solution was pre-incubated for 2 hrs.</div></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=211235&oldid=prevSlecat: /* Transformation */2012-09-26T19:11:22Z<p><span class="autocomment">Transformation</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Splead 300 ul of LB&DH5&alpha; solution to LBK.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Splead 300 ul of LB&DH5&alpha; solution to LBK.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated for 19 hrs.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated for 19 hrs.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div></p> <del class="diffchange diffchange-inline"> </del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div></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">*K346007(Ag43)</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: #eee; color:black; font-size: smaller;"><div>==Plasmid extraction==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Plasmid extraction==</div></td></tr>
<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>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=211207&oldid=prevSlecat: /* Glycerol stock */2012-09-26T19:10:43Z<p><span class="autocomment">Glycerol stock</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Glycerol stock===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Glycerol stock===</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">glycerol </del>stocks of dT,RBS,pT7 and pLacI-RBS-Ag43.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Glycerol </ins>stocks of dT,RBS,pT7 and pLacI-RBS-Ag43.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Parts written above were incubated in 1 ml of LBA(dT,RBS,pT7) and LBC(pLacI-RBS-Ag43) for 16 hrs 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Parts written above were incubated in 1 ml of LBA(dT,RBS,pT7) and LBC(pLacI-RBS-Ag43) for 16 hrs 30 min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add glycerol and Freeze at -80C</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Add glycerol and Freeze at -80C</div></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=207579&oldid=prevSlecat at 17:53, 26 September 20122012-09-26T17:53:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==July 6th==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==July 6th==</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 class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>===Incubation for <del class="diffchange diffchange-inline">mini-prep </del>and glycerol stock===</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>===Incubation for <ins class="diffchange diffchange-inline">plasmid extraction </ins>and glycerol stock===</div></td></tr>
<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>Incubated in LBA(dT,RBS,pT7) and LBC(pLacI-RBS-Ag43) solution.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Incubated in LBA(dT,RBS,pT7) and LBC(pLacI-RBS-Ag43) solution.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Picked up two colonies from each plate.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Picked up two colonies from each plate.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#First colony was re-suspended in 1 ml LB (A and C respectively) for glycerol stock. Second colony was re-suspended in 2 ml LB(A and C respectively) for <del class="diffchange diffchange-inline">mini-prep</del>. </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#First colony was re-suspended in 1 ml LB (A and C respectively) for glycerol stock. Second colony was re-suspended in 2 ml LB(A and C respectively) for <ins class="diffchange diffchange-inline">plasmid extraction</ins>. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated for 16 hrs.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated for 16 hrs.</div></td></tr>
<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>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==July 7th==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==July 7th==</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 class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>===Incubation of <del class="diffchange diffchange-inline">mini-prep </del>and glycerol stock===</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>===Incubation of <ins class="diffchange diffchange-inline">plasmid extraction </ins>and glycerol stock===</div></td></tr>
<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>Incubation of (Ag43) in LBC liquid medium.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Incubation of (Ag43) in LBC liquid medium.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Plasmid extraction===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Plasmid extraction===</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">mini-prep </del>of dT,RBS,pT7 and pLacI-RBS-Ag43. We used FastGene Plasmid Mini Kit(Nippon Gene)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Plasmid extraction </ins>of dT,RBS,pT7 and pLacI-RBS-Ag43. We used FastGene Plasmid Mini Kit(Nippon Gene)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#We got 50 ul of DNA solutions.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#We got 50 ul of DNA solutions.</div></td></tr>
<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>
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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>[[Image:120707_I719005_B0034_B0015_K542009_mini-prep_umeuchi.jpg|thumb|Erectrophoresis result]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:120707_I719005_B0034_B0015_K542009_mini-prep_umeuchi.jpg|thumb|Erectrophoresis result]]</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Electrophoresis to estimate the concentration of <del class="diffchange diffchange-inline">mini-prep </del>products(dT,RBS,pT7 and pLacI-RBS-Ag43).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Electrophoresis to estimate the concentration of <ins class="diffchange diffchange-inline">plasmid extraction </ins>products(dT,RBS,pT7 and pLacI-RBS-Ag43).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Used 1% agarose gel.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Used 1% agarose gel.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added EtBr to TBE buffer and soaked the gel by applying current.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added EtBr to TBE buffer and soaked the gel by applying current.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Ran '''1.2 ul of DNA solutions (1 ul is <del class="diffchange diffchange-inline">mini-prep </del>product and 0.2 ul is Loading Dye)''' for 35 min.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Ran '''1.2 ul of DNA solutions (1 ul is <ins class="diffchange diffchange-inline">plasmid extraction </ins>product and 0.2 ul is Loading Dye)''' for 35 min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Took a photograph of 1% agarose gel.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Took a photograph of 1% agarose gel.</div></td></tr>
<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>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Plasmid extraction==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Plasmid extraction==</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">mini-prep </del>for Liquid culture product of K346007(Ag43)</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">Plasmid extraction </ins>for Liquid culture product of K346007(Ag43)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Used FastGene Plasmid Mini Kit(Nippon Genetics)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Used FastGene Plasmid Mini Kit(Nippon Genetics)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Elutioned in 50 ul</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Elutioned in 50 ul</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Electrophoresis===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Electrophoresis===</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div>[[image:HokkaidoU2012 120708 K346007 miniprep and pt7 rbs.jpg|thumb|<del class="diffchange diffchange-inline">mini-prep </del>result (With ligation result of pT7+RBS+pSB1K3)]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>[[image:HokkaidoU2012 120708 K346007 miniprep and pt7 rbs.jpg|thumb|<ins class="diffchange diffchange-inline">plasmid extraction </ins>result (With ligation result of pT7+RBS+pSB1K3)]]</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Electrophoresis for <del class="diffchange diffchange-inline">mini-prep </del>product(Ag43).</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Electrophoresis for <ins class="diffchange diffchange-inline">plasmid extraction </ins>product(Ag43).</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Prepared 1% Agalose gel and added EtBr then pre-migration for 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Prepared 1% Agalose gel and added EtBr then pre-migration for 30 min.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#1 ul 1kb ladder, 1.2 ul <del class="diffchange diffchange-inline">mini-prep </del>product(1 ul is DNA solution and 0.2 ul is loading dye) added then migtrated.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#1 ul 1kb ladder, 1.2 ul <ins class="diffchange diffchange-inline">plasmid extraction </ins>product(1 ul is DNA solution and 0.2 ul is loading dye) added then migtrated.</div></td></tr>
<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>===Glycerol stock===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Glycerol stock===</div></td></tr>
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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>Digested Ag43 and dT in solution by recipes Written below.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Digested Ag43 and dT in solution by recipes Written below.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>'''Insert DNA required too much weight and volume'''(volume was calculated from concentration of <del class="diffchange diffchange-inline">DNA mini-prep</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>'''Insert DNA required too much weight and volume'''(volume was calculated from concentration of <ins class="diffchange diffchange-inline">plasmid extraction</ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>product)from our calculation. There are no insurance of succession of digestion. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>product)from our calculation. There are no insurance of succession of digestion. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*Ag43(Insert)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*Ag43(Insert)</div></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=207465&oldid=prevSlecat: /* Incubation of mini-prep and glycerol stock */2012-09-26T17:50:47Z<p><span class="autocomment">Incubation of mini-prep and glycerol stock</span></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 17:50, 26 September 2012</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>Incubation of (Ag43) in LBC liquid medium.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Incubation of (Ag43) in LBC liquid medium.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">#</del>Two colonies were re-suspended in 2 ml LBC. We incubated them at 38C.<br /></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><br /></ins>Two colonies were re-suspended in 2 ml LBC. We incubated them at 38C.<br /></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: #eee; color:black; font-size: smaller;"><div>'''One of the tubes was incubated for 8 hrs. It's for glycerol stock.'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''One of the tubes was incubated for 8 hrs. It's for glycerol stock.'''</div></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=207398&oldid=prevSlecat: /* Transformation */2012-09-26T17:48:58Z<p><span class="autocomment">Transformation</span></p>
<table style="background-color: white; color:black;">
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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 17:48, 26 September 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transformation===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Transformation===</div></td></tr>
<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: #ffa; color:black; font-size: smaller;"><div>'''K346007(Ag43) failed to <del class="diffchange diffchange-inline">produce colonies </del>on LBC plate.'''</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>'''K346007(Ag43) failed to <ins class="diffchange diffchange-inline">grow </ins>on LBC plate.'''</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: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=192338&oldid=prevSlecat at 10:53, 26 September 20122012-09-26T10:53:54Z<p></p>
<table style="background-color: white; color:black;">
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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 10:53, 26 September 2012</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 3 ul of NaoAc, 1.5 ul of glycogen and 75 ul of 100% ethanol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 3 ul of NaoAc, 1.5 ul of glycogen and 75 ul of 100% ethanol.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 14000 rpm, 30 min at 4C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 14000 rpm, 30 min at 4C.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">Remove </del>supernatant and added 220ul of 70% ethanol.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Removed </ins>supernatant and added 220ul of 70% ethanol.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 15 min at 4C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 15 min at 4C.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Removed supernatant and air dried at room temperature. Dissolved in 10 ul of DW.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Removed supernatant and air dried at room temperature. Dissolved in 10 ul of DW.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 5 ul of NaoAc , 1.5 ul of glycogen and 125 ul of 100% ethanol to 50 ul DNA solutions.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 5 ul of NaoAc , 1.5 ul of glycogen and 125 ul of 100% ethanol to 50 ul DNA solutions.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 10min at 4C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 10min at 4C.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">Remove </del>supernatant and added 220 ul of 70% ethanol.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Removed </ins>supernatant and added 220 ul of 70% ethanol.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 5 min at 4C.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Centrifuged at 15000 rpm, 5 min at 4C.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">Remove </del>supernatant and air drying at room temperature then added 10 ul of DW.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Removed </ins>supernatant and air drying at room temperature then added 10 ul of DW.</div></td></tr>
<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>===Ligation===</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>===Ligation===</div></td></tr>
</table>Slecathttp://2012.igem.org/wiki/index.php?title=Team:HokkaidoU_Japan/Notebook/aggregation_Week_1&diff=191955&oldid=prevSlecat at 10:38, 26 September 20122012-09-26T10:38:22Z<p></p>
<table style="background-color: white; color:black;">
<col class='diff-marker' />
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<col class='diff-marker' />
<col class='diff-content' />
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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 10:38, 26 September 2012</td>
</tr><tr><td colspan="2" class="diff-lineno">Line 44:</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Transformation of K346007(Ag43) to DH5&alpha;.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Transformation of K346007(Ag43) to DH5&alpha;.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Mixed 1 ul DNA solution with DH5&alpha; competent cells and incubated on ice for 30 min.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#To gain chrolamphenicol resistance solution was pre-<del class="diffchange diffchange-inline">cultivated </del>for 2 hrs.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#To gain chrolamphenicol resistance solution was pre-<ins class="diffchange diffchange-inline">incubated </ins>for 2 hrs.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated on LBC for 21 hrs.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Incubated on LBC for 21 hrs.</div></td></tr>
<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 colspan="2" class="diff-lineno">Line 257:</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Stood on ice for 30 min.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Stood on ice for 30 min.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 600 ul of LB to transformed DH5&alpha; solution.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added 600 ul of LB to transformed DH5&alpha; solution.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Pre-<del class="diffchange diffchange-inline">cultivate </del>for 2 hrs at 37C.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Pre-<ins class="diffchange diffchange-inline">incubated </ins>for 2 hrs at 37C.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Splead 300 ul of LB&DH5&alpha; solution to LBK.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Splead 300 ul of LB&DH5&alpha; solution to LBK.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">Cultivated</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Incubated for 19 hrs.</ins></div></td></tr>
<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>*K346007(Ag43)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>*K346007(Ag43)</div></td></tr>
<tr><td colspan="2" class="diff-lineno">Line 279:</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>Made glycerol stock of K346007 (Ag43).</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Made glycerol stock of K346007 (Ag43).</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#Parts written above were <del class="diffchange diffchange-inline">cultivated </del>in LBC.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#Parts written above were <ins class="diffchange diffchange-inline">incubated </ins>in LBC.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added glycerol and freezed at -80C</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Added glycerol and freezed at -80C</div></td></tr>
<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>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#From 700 solution(100 ul is DH5&alpha; and 600 ul is LB), 100 ul add to 900 ul of LB(x10 solution)</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#From 700 solution(100 ul is DH5&alpha; and 600 ul is LB), 100 ul add to 900 ul of LB(x10 solution)</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Spread 300 ul from 600(700-100) ul and 1000 ul of LB&DH5&alpha; solution to each LBA plates.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>#Spread 300 ul from 600(700-100) ul and 1000 ul of LB&DH5&alpha; solution to each LBA plates.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>#<del class="diffchange diffchange-inline">Cultivated</del>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>#<ins class="diffchange diffchange-inline">Incubated for 19 hrs</ins>.</div></td></tr>
<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></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></div></div></div></td></tr>
</table>Slecat