http://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&feed=atom&action=historyTeam:Paris Bettencourt/Project/Interlab Study - Revision history2024-03-29T07:09:50ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=403738&oldid=prevPsatin at 16:05, 7 December 20142014-12-07T16:05:31Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br></br></br><h5>Devices</h5></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br></br></br><h5>Devices</h5></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div class=text><p>We built 3 devices (Fig. 1, Fig. 2, Fig. 3) according to <a href"https://2014.igem.org/Tracks/Measurement/Interlab_study">Interlab study instructions</a>. First designed in software (Geneious v. 7.0.6). Then made in our wet lab using the Biobrick Distribution Kits and confirmed by <del class="diffchange diffchange-inline">eletrophoresis </del>gel analytic digestion as well as sequencing. Finally, stocked as a glycerol stock and used for the <del class="diffchange diffchange-inline">measurments </del>of GFP expression. </p></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div class=text><p>We built 3 devices (Fig. 1, Fig. 2, Fig. 3) according to <a href"https://2014.igem.org/Tracks/Measurement/Interlab_study">Interlab study instructions</a>. First designed in software (Geneious v. 7.0.6). Then made in our wet lab using the Biobrick Distribution Kits and confirmed by <ins class="diffchange diffchange-inline">electrophoresis </ins>gel analytic digestion as well as sequencing. Finally, stocked as a glycerol stock and used for the <ins class="diffchange diffchange-inline">measurements </ins>of GFP expression. </p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td></tr>
</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=403737&oldid=prevPsatin at 15:16, 7 December 20142014-12-07T15:16:30Z<p></p>
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</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=403736&oldid=prevPsatin at 15:16, 7 December 20142014-12-07T15:16:09Z<p></p>
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</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=391413&oldid=prevImarcus at 02:47, 18 October 20142014-10-18T02:47:06Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></br></br></br><h5>Results</h5></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></br></br></br><h5>Results</h5></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>Here we present the result of the measurments. The measument has been realised in a comparable growth conditions for all Devices (Fig. 4). We can clearly see that the highest GFP expression levels occures under <a href="http://parts.igem.org/Part:BBa_J23101"> BBa_J23101</a> Anderson's strong promoter in a high copy plasmid sPB1C3 (Device 2: <a href="http://parts.igem.org/Part:BBa_K1403000">BBa_K1403000</a>). The lowest GFP levels occures under very weak Anderson's promoter - mutated <a href="http://parts.igem.org/Part:BBa_J23101"> J23115</a> (Device 3: <a href="http://parts.igem.org/Part:BBa_K1403001">BBa_K1403001</a>)as can be seen in the Fig. 5 & Fig. 6 </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>Here we present the result of the measurments. The measument has been realised in a comparable growth conditions for all Devices (Fig. 4). We can clearly see that the highest GFP expression levels occures under <a href="http://parts.igem.org/Part:BBa_J23101"> BBa_J23101</a> Anderson's strong promoter in a high copy plasmid sPB1C3 (Device 2: <a href="http://parts.igem.org/Part:BBa_K1403000">BBa_K1403000</a>). The lowest GFP levels occures under very weak Anderson's promoter - mutated <a href="http://parts.igem.org/Part:BBa_J23101"> J23115</a> (Device 3: <a href="http://parts.igem.org/Part:BBa_K1403001">BBa_K1403001</a>) as can be seen in the Fig. 5 & Fig. 6 </p></div></td></tr>
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</table>Imarcushttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=391238&oldid=prevImarcus at 02:45, 18 October 20142014-10-18T02:45:48Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li>BBa_I20260: Plate 2, Well 17F</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li>BBa_I20260: Plate 2, Well 17F</li></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p> We followed <a href=kit>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick Plate 2, Well 17F (2012) and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock transformed the <i>E. coli</i></a> colonies that grew in the selective Kanymycin were grown in liquid media and made into <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stocks</a> <del class="diffchange diffchange-inline">and </del>labelled G.22.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p> We followed <a href=kit>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick Plate 2, Well 17F (2012) and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock transformed the <i>E. coli</i></a> colonies that grew in the selective Kanymycin were grown in liquid media and made into <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stocks</a> labelled G.22.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id=divimage class=image><div class=separation2></div><img src="https://static.igem.org/mediawiki/2014/0/02/Device_2.png"><p class=definition><b>Figure 2. Device 2.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id=divimage class=image><div class=separation2></div><img src="https://static.igem.org/mediawiki/2014/0/02/Device_2.png"><p class=definition><b>Figure 2. Device 2.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> Complete with H2O</li></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> Complete with H2O</li></ul></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (Final volume of 50 uL)</br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> (Final volume of 50 uL)</br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> We made an eletrophoresis gel to check the fragments (the bands at around 876 bp for GFP and 2100 bp for the promoter + backbone)and then <del class="diffchange diffchange-inline">extract </del>BBa_E0240 with <del class="diffchange diffchange-inline">Gel </del>extraction kit. For the plasmid with the promoter we used a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot5">PCR purification kit.</a></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> We made an eletrophoresis gel to check the fragments (the bands at around 876 bp for GFP and 2100 bp for the promoter + backbone) and then <ins class="diffchange diffchange-inline">extracted </ins>BBa_E0240 with <ins class="diffchange diffchange-inline">a gel </ins>extraction kit. For the plasmid with the promoter we used a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot5">PCR purification kit.</a></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> We introduced the GFP fragment to the Plasmid + backbone through ligation of the sticky ends SpeI and XbeI. Quantified DNA in two parts with nanodrop. The amount of vector:insert has been calculated with Promega calculator. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> We introduced the GFP fragment to the Plasmid + backbone through ligation of the sticky ends SpeI and XbeI. Quantified DNA in two parts with nanodrop. The amount of vector:insert has been calculated with Promega calculator. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Incubate at 22°C for 1h</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li>Incubate at 22°C for 1h</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> 16°C overnight</li></ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> 16°C overnight</li></ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> We transformed the ligation product following <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock transformation of <i>E.coli</i></a>.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> We transformed the ligation product following <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock transformation of <i>E. coli</i></a>.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> We made liquid <del class="diffchange diffchange-inline">cukltures </del>of single colonies with the appropriate antibiotic and the next day we prepared a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stock </a>.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> We made liquid <ins class="diffchange diffchange-inline">cultures </ins>of single colonies with the appropriate antibiotic and the next day we prepared a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stock</a>.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image><img src="https://static.igem.org/mediawiki/2014/d/d9/Device_3.png"><p class=definition><b>Figure 3. Device 3.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image><img src="https://static.igem.org/mediawiki/2014/d/d9/Device_3.png"><p class=definition><b>Figure 3. Device 3.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Our part in the registry: <a href="http://parts.igem.org/Part:BBa_K1403001">BBa_K1403001</a></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>Our part in the registry: <a href="http://parts.igem.org/Part:BBa_K1403001">BBa_K1403001</a></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p><i>*BBa_J23115 was cloned using BBa_K823012 and therefore should have 2 <del class="diffchange diffchange-inline">missmatched </del>basepairs.</i></p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p><i>*BBa_J23115 was cloned using BBa_K823012 and therefore should have 2 <ins class="diffchange diffchange-inline">mismatched </ins>basepairs.</i></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><u>BBa_J23115 + BBa_E0240</u> (B0032-E0040-B0010-B0012), in the <u>pSB1C3</u> vector.</br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><u>BBa_J23115 + BBa_E0240</u> (B0032-E0040-B0010-B0012), in the <u>pSB1C3</u> vector.</br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Selection marker : Chloramphenicol</br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Selection marker : Chloramphenicol</br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Promoter expected sequence : tttatagctagctcag<u>t</u>cct<u>a</u>ggtacaatgctagc (<del class="diffchange diffchange-inline">missmatched </del>basepairs compared to real BBa_J23115 are underlined)</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Promoter expected sequence : tttatagctagctcag<u>t</u>cct<u>a</u>ggtacaatgctagc (<ins class="diffchange diffchange-inline">mismatched </ins>basepairs compared to real BBa_J23115 are underlined)</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><u>2014 Biobrick Kit locations</u></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><u>2014 Biobrick Kit locations</u></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li> BBa_K823012 (BBa_J23115 in pSB1C3): Plate 1, Well 22I</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li> BBa_K823012 (BBa_J23115 in pSB1C3): Plate 1, Well 22I</li></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>In order to prepare the third device we <del class="diffchange diffchange-inline">proceed </del>exactly in the same way as for the Device 2, except we used BBa_K823012 instead of BBa_K823005</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>In order to prepare the third device we <ins class="diffchange diffchange-inline">proceeded </ins>exactly in the same way as for the Device 2, except we used BBa_K823012 instead of BBa_K823005</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
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</table>Imarcushttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=390593&oldid=prevImarcus at 02:41, 18 October 20142014-10-18T02:41:02Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class=text1><i>"The goal of the interlab study is to obtain fluorescence data for three specific genetic devices expressing GFP from iGEM teams around the world. "</i></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p class=text1><i>"The goal of the interlab study is to obtain fluorescence data for three specific genetic devices expressing GFP from iGEM teams around the world. "</i></p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h5>Motivation</h5></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h5>Motivation</h5></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p class=text1>iGEM Paris Bettencourt team supported this initiative with pleasure. We strongly believe that this study will help to define standards for BioBrick constructs and measure the difference between different labs and measurement methods. It was also a great training for inexperienced synthetic biologist <del class="diffchange diffchange-inline">of </del>the team <del class="diffchange diffchange-inline">which allowed as to train transformation and ligation techniques</del>. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p class=text1>iGEM Paris Bettencourt team supported this initiative with pleasure. We strongly believe that this study will help to define standards for BioBrick constructs and measure the difference between different labs and measurement methods. It was also a great training for <ins class="diffchange diffchange-inline">transformation and ligation techniques for the </ins>inexperienced synthetic biologist <ins class="diffchange diffchange-inline">on </ins>the team.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td 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> </br></br></br><h5>Devices</h5></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br></br></br><h5>Devices</h5></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div class=text><p>We built 3 devices (Fig. 1, Fig. 2, Fig. 3) according to <a href"https://2014.igem.org/Tracks/Measurement/Interlab_study">Interlab study instructions</a>. First designed in <del class="diffchange diffchange-inline">a </del>software (Geneious v. 7.0.6). Then <del class="diffchange diffchange-inline">realised </del>in our wet lab <del class="diffchange diffchange-inline">out of </del>Biobrick Distribution Kits<del class="diffchange diffchange-inline">. Confirmed </del>by eletrophoresis gel analytic <del class="diffchange diffchange-inline">dgestion </del>as well as sequencing. Finally, stocked as a glycerol stock and used for the measurments of GFP expression. </p></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div class=text><p>We built 3 devices (Fig. 1, Fig. 2, Fig. 3) according to <a href"https://2014.igem.org/Tracks/Measurement/Interlab_study">Interlab study instructions</a>. First designed in <ins class="diffchange diffchange-inline"> </ins>software (Geneious v. 7.0.6). Then <ins class="diffchange diffchange-inline">made </ins>in our wet lab <ins class="diffchange diffchange-inline">using the </ins>Biobrick Distribution Kits <ins class="diffchange diffchange-inline">and confirmed </ins>by eletrophoresis gel analytic <ins class="diffchange diffchange-inline">digestion </ins>as well as sequencing. Finally, stocked as a glycerol stock and used for the measurments of GFP expression. </p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li>BBa_I20260: Plate 2, Well 17F</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li>BBa_I20260: Plate 2, Well 17F</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p> We followed <a href=kit>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick Plate 2, Well 17F (2012) and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock <del class="diffchange diffchange-inline">transformation of </del><i>E.coli</i></a><del class="diffchange diffchange-inline">Colonies </del>that grew in <del class="diffchange diffchange-inline">a </del>selective Kanymycin were grown and <del class="diffchange diffchange-inline">stocked in the </del><a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol <del class="diffchange diffchange-inline">stock </del></a> and <del class="diffchange diffchange-inline">labbeled it </del>G.22.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p> We followed <a href=kit>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick Plate 2, Well 17F (2012) and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock <ins class="diffchange diffchange-inline">transformed the </ins><i>E. coli</i></a> <ins class="diffchange diffchange-inline">colonies </ins>that grew in <ins class="diffchange diffchange-inline">the </ins>selective Kanymycin were grown <ins class="diffchange diffchange-inline">in liquid media </ins>and <ins class="diffchange diffchange-inline">made into </ins><a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol <ins class="diffchange diffchange-inline">stocks</ins></a> and <ins class="diffchange diffchange-inline">labelled </ins>G.22.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id=divimage class=image><div class=separation2></div><img src="https://static.igem.org/mediawiki/2014/0/02/Device_2.png"><p class=definition><b>Figure 2. Device 2.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div id=divimage class=image><div class=separation2></div><img src="https://static.igem.org/mediawiki/2014/0/02/Device_2.png"><p class=definition><b>Figure 2. Device 2.</b>Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> BBa_E0240 (in pSB1C3): Plate 2, Well 24B</li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li> BBa_E0240 (in pSB1C3): Plate 2, Well 24B</li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </ul></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <p> We followed the <a>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick BBa_K823005 and BBa_E0240 and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock <del class="diffchange diffchange-inline">transformation of </del><i>E.coli</i></a></br>The colonies that <del class="diffchange diffchange-inline">survided </del>after selection in choloamphenicol <del class="diffchange diffchange-inline">werecultured </del>overnight. We used 750uL of the liquid cultures for a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stock </a>. <del class="diffchange diffchange-inline">We used </del>remaining 4<del class="diffchange diffchange-inline">,</del>25 mL to make <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4"> minipreps <del class="diffchange diffchange-inline"> </del></a>. <del class="diffchange diffchange-inline">we </del>measured DNA content with the nanodrop.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <p> We followed the <a>iGEM Distribution Kit instructions </a> to extract DNA from the Biobrick BBa_K823005 and BBa_E0240 and then <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1"> Heat Shock <ins class="diffchange diffchange-inline">transformed </ins><i>E. coli</i></a></br>The colonies that <ins class="diffchange diffchange-inline">survived </ins>after selection in choloamphenicol <ins class="diffchange diffchange-inline">were cultured </ins>overnight. We used 750uL of the liquid cultures for a <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">glycerol stock </a>. <ins class="diffchange diffchange-inline">The </ins>remaining 4<ins class="diffchange diffchange-inline">.</ins>25 mL <ins class="diffchange diffchange-inline">were used </ins>to make <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4"> minipreps </a>. <ins class="diffchange diffchange-inline">We </ins>measured DNA content with the nanodrop.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></br></br><u>Digestion analysis</u>:</br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></br></br><u>Digestion analysis</u>:</br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li> 5 ug plasmid </li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <ul><li> 5 ug plasmid </li></div></td></tr>
</table>Imarcushttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=383266&oldid=prevPsatin at 01:36, 18 October 20142014-10-18T01:36:42Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> width : <del class="diffchange diffchange-inline">100</del>%;</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> width : <ins class="diffchange diffchange-inline">75</ins>%;</div></td></tr>
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</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=383184&oldid=prevPsatin at 01:36, 18 October 20142014-10-18T01:36:01Z<p></p>
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<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;"> margin-left : 12.5%;</del></div></td><td colspan="2"> </td></tr>
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</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=383142&oldid=prevPsatin at 01:35, 18 October 20142014-10-18T01:35:40Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> top : 800px;</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> top : 800px;</div></td></tr>
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</table>Psatinhttp://2014.igem.org/wiki/index.php?title=Team:Paris_Bettencourt/Project/Interlab_Study&diff=370083&oldid=prevPsatin: Undo revision 370038 by Psatin (talk)2014-10-17T23:44:21Z<p>Undo revision 370038 by <a href="/Special:Contributions/Psatin" title="Special:Contributions/Psatin">Psatin</a> (<a href="/wiki/index.php?title=User_talk:Psatin&action=edit&redlink=1" class="new" title="User talk:Psatin (page does not exist)">talk</a>)</p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=image id=firstimg><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </br><h6>Device 1</h6></br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> <div class=image id=<del class="diffchange diffchange-inline">secondimg</del>><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></div></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> <div class=image id=<ins class="diffchange diffchange-inline">firstimg</ins>><img src="https://static.igem.org/mediawiki/2014/9/91/Device_1.png"><p class=definition><b>Figure 1. Device 1.</b> Geneious version 7.0.6 created by Biomatters. Available from http://www.geneious.com/</p></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: #eee; color:black; font-size: smaller;"><div> <div class=text><p><u>BBa_I20260</u> (J23101-B0032-E0040-B0010-B0012) in the <u>pSB3K3</u> vector.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <div class=text><p><u>BBa_I20260</u> (J23101-B0032-E0040-B0010-B0012) in the <u>pSB3K3</u> vector.</p></div></td></tr>
</table>Psatin