http://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&feed=atom&action=historyTeam:CIDEB-UANL Mexico/labwork conclusions - Revision history2024-03-29T11:57:53ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27909&oldid=prevFernandaPuente at 03:59, 21 June 20142014-06-21T03:59:21Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">Also, the tests</del>, at first, where performed in test tubes. After obtaining an aroma result, the team did the same experiment in Petri dishes. It was appreciated that the odor was more intense in Petri dishes than in test tubes.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">As an extra</ins>, at first, <ins class="diffchange diffchange-inline">the experiments </ins>where performed in test tubes. After obtaining an aroma result, the team did the same experiment in Petri dishes. It was appreciated that the odor was more intense in Petri dishes than in test tubes.</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>
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</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27893&oldid=prevFernandaPuente at 03:58, 21 June 20142014-06-21T03:58:32Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>It was appreciated that the odor was more intense in Petri dishes than in test tubes.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">Also, the tests, at first, where performed in test tubes. After obtaining an aroma result, the team did the same experiment in Petri dishes. </ins>It was appreciated that the odor was more intense in Petri dishes than in test tubes.</p></div></td></tr>
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</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27858&oldid=prevFernandaPuente at 03:55, 21 June 20142014-06-21T03:55:29Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubated at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </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 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>It was appreciated that the odor was more intense in Petri dishes than in test tubes.</p></ins></div></td></tr>
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</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27846&oldid=prevFernandaPuente at 03:54, 21 June 20142014-06-21T03:54:13Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. <del class="diffchange diffchange-inline"> </del>In the case of the plaque that had the odor even if it was <del class="diffchange diffchange-inline">incubate </del>at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor. If the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was <ins class="diffchange diffchange-inline">incubated </ins>at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of the WinterGreen enzyme by saturating it. </p></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="text-align: right;"><a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_conclusions#"><font color="blue">Return to the Top</font></a></p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="text-align: right;"><a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_conclusions#"><font color="blue">Return to the Top</font></a></p></div></div></td></tr>
</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27474&oldid=prevFernandaPuente at 03:24, 21 June 20142014-06-21T03:24:08Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Aroma module</b><p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor <del class="diffchange diffchange-inline">and if </del>the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch <del class="diffchange diffchange-inline">actually </del>worked at 35°C. In the case of the plaque that had the odor even if it was incubate at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of <del class="diffchange diffchange-inline">winter green </del>enzyme by saturating it. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor<ins class="diffchange diffchange-inline">. If </ins>the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch worked at 35°C. In the case of the plaque that had the odor even if it was incubate at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of <ins class="diffchange diffchange-inline">the WinterGreen </ins>enzyme by saturating it. </p></div></td></tr>
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</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27450&oldid=prevFernandaPuente at 03:22, 21 June 20142014-06-21T03:22:23Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As it was reported in the patent, the nhaS gene gave certain resistance to salt but there was not any exact percentage of the resistance. There were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium <del class="diffchange diffchange-inline">with only </del>NaCl it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As it was reported in the patent, the nhaS gene gave certain resistance to salt but there was not any exact percentage of the resistance. There were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium <ins class="diffchange diffchange-inline">containing </ins>NaCl <ins class="diffchange diffchange-inline">only </ins>it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</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>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></div></td></tr>
</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27441&oldid=prevFernandaPuente at 03:21, 21 June 20142014-06-21T03:21:26Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As it was reported, the nhaS gene gave certain resistance to salt but there was not any exact percentage of the resistance. There were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium with only NaCl it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As it was reported <ins class="diffchange diffchange-inline">in the patent</ins>, the nhaS gene gave certain resistance to salt but there was not any exact percentage of the resistance. There were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium with only NaCl it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</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>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></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"> Give </del>resistance to salty environments up to 15% of NaCl concentration</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">●Give </ins>resistance to salty environments up to 15% of NaCl concentration</p></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"> Captures </del>ion sodium ions</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">●Captures </ins>ion sodium ions</p></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"> Regulates </del>the pH of the cell (this is the reason of the resistance it gives)</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p><ins class="diffchange diffchange-inline">●Regulates </ins>the pH of the cell (this is the reason of the resistance it gives)</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the case of the E.CARU project, those characteristics where used with the purpose of remove sodium ions in order to desalinize water, but nhaS can be used in many different forms and in other aspects of biotechnology.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the case of the E.CARU project, those characteristics where used with the purpose of remove sodium ions in order to desalinize water, but nhaS can be used in many different forms and in other aspects of biotechnology.</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>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27405&oldid=prevFernandaPuente at 03:18, 21 June 20142014-06-21T03:18:47Z<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 03:18, 21 June 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Capture module</b></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Capture module</b></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>After all the experiments and results interpretations, it was concluded that even if the ligation of nhaS and RFP with pSB1C3 came out with two types of bacteria (red and white), both of them had the nhaS gene functioning. The reasons of why there were red and white colonies, first, is that the Petri <del class="diffchange diffchange-inline">dish wherethe </del>transformed bacteria were inoculated with the gene, <del class="diffchange diffchange-inline">was </del>not covered <del class="diffchange diffchange-inline">by </del>aluminum. This meant that the UV promotor was activated with normal light and activated the production of nhaS and RFP. The second reason is that during the purification process, the RBS and the first 50 nucleotides of the RFP region mutated because of the exposure to UV irradiation, a necessary step in the process. Because of this, bacteria transformed with the ligation of mutated fragments would not express the RFP gene, and therefore they appeared white.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>After all the experiments and results interpretations, it was concluded that even if the ligation of nhaS and RFP with pSB1C3 came out with two types of bacteria (red and white), both of them had the nhaS gene functioning.<ins class="diffchange diffchange-inline"></p> <p></ins>The reasons of why there were red and white colonies, first, is that the Petri <ins class="diffchange diffchange-inline">dishes where the </ins>transformed bacteria were inoculated with the gene, <ins class="diffchange diffchange-inline">were </ins>not covered <ins class="diffchange diffchange-inline">with </ins>aluminum. This meant that the UV promotor was activated with normal light and activated the production of nhaS and RFP. The second reason is that during the purification process, the RBS and the first 50 nucleotides of the RFP region mutated because of the exposure to UV irradiation, a necessary step in the process. Because of this, bacteria transformed with the ligation of mutated fragments would not express the RFP gene, and therefore they appeared white.</p></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</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>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=27381&oldid=prevFernandaPuente at 03:16, 21 June 20142014-06-21T03:16:06Z<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 03:16, 21 June 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Capture module</b></p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p><b>Capture module</b></p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>After all the experiments and <del class="diffchange diffchange-inline">interpretations of </del>results, it <del class="diffchange diffchange-inline">be </del>concluded that even if the ligation of nhaS<del class="diffchange diffchange-inline">+</del>RFP with pSB1C3 <del class="diffchange diffchange-inline">gave </del>two types of bacteria (red and white), both of them had the nhaS gene functioning. The reasons of why there <del class="diffchange diffchange-inline">was </del>red and white colonies, first, is that the Petri dish <del class="diffchange diffchange-inline">where </del>were inoculated <del class="diffchange diffchange-inline">the transformed bacteria </del>with the gene, was not covered by aluminum<del class="diffchange diffchange-inline">, so </del>the UV promotor was activated with normal light and activated the production of nhaS and RFP. The second reason is that during the purification process the RBS and the first 50 nucleotides of <del class="diffchange diffchange-inline">RPF </del>region mutated because of the exposure to UV irradiation, a necessary step in the process. <del class="diffchange diffchange-inline">Then</del>, bacteria transformed with the ligation of mutated fragments<del class="diffchange diffchange-inline">, </del>would not express the RFP gene, therefore they <del class="diffchange diffchange-inline">were </del>white.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>After all the experiments and results <ins class="diffchange diffchange-inline">interpretations</ins>, it <ins class="diffchange diffchange-inline">was </ins>concluded that even if the ligation of nhaS <ins class="diffchange diffchange-inline">and </ins>RFP with pSB1C3 <ins class="diffchange diffchange-inline">came out with </ins>two types of bacteria (red and white), both of them had the nhaS gene functioning. The reasons of why there <ins class="diffchange diffchange-inline">were </ins>red and white colonies, first, is that the Petri dish <ins class="diffchange diffchange-inline">wherethe transformed bacteria </ins>were inoculated with the gene, was not covered by aluminum<ins class="diffchange diffchange-inline">. This meant that </ins>the UV promotor was activated with normal light and activated the production of nhaS and RFP. The second reason is that during the purification process<ins class="diffchange diffchange-inline">, </ins>the RBS and the first 50 nucleotides of <ins class="diffchange diffchange-inline">the RFP </ins>region mutated because of the exposure to UV irradiation, a necessary step in the process. <ins class="diffchange diffchange-inline">Because of this</ins>, bacteria transformed with the ligation of mutated fragments would not express the RFP gene, <ins class="diffchange diffchange-inline">and </ins>therefore they <ins class="diffchange diffchange-inline">appeared </ins>white.</p></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria<del class="diffchange diffchange-inline">, </del>and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>The mutation of the RBS before RFP causing white bacteria and the functionality presence of nhaS in both types of colonies was proved in the sequencing and in the experiments of viability in salt.</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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p>As it was reported <del class="diffchange diffchange-inline">that </del>the <del class="diffchange diffchange-inline">gen </del>nhaS <del class="diffchange diffchange-inline"> </del>gave certain resistance to salt but there was not any exact percentage of the resistance<del class="diffchange diffchange-inline">, there </del>were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium with only NaCl it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><p>As it was reported<ins class="diffchange diffchange-inline">, </ins>the nhaS <ins class="diffchange diffchange-inline">gene </ins>gave certain resistance to salt but there was not any exact percentage of the resistance<ins class="diffchange diffchange-inline">. There </ins>were performed diverse experiments in order to know the maximum salt concentration bacteria transformed with the gene would survive. The concentrations where the bacteria survived were from 1% up to 15% of NaCl in the medium. It can be concluded that the bacteria survived in different NaCl concentrations up to 15%, while not transformed bacteria can survive in a 1% NaCl medium, but not in higher NaCl concentrated mediums. Another conclusion obtained from the experiments is that the bacteria with NhaS can survive in a high NaCl concnetrated medium only if it has its corresponding nutrients, because in a medium with only NaCl it dies. We already know that the bacteria survives in a high NaCl medium, but the experiments that prove whether the bacteria captures sodium ions or not will be performed the weekend of June 21th and 22th. The results will be shown in the Jamboree presentation.</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>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In summary, nhaS is a gene that produce a multitask protein with a lot of advantages, not only because it is pretty little and do not represent a large genetic charge to the bacteria, but also it gives many abilities to it:</p></div></td></tr>
</table>FernandaPuentehttp://2014hs.igem.org/wiki/index.php?title=Team:CIDEB-UANL_Mexico/labwork_conclusions&diff=26102&oldid=prevMarea at 01:28, 21 June 20142014-06-21T01:28:25Z<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 01:28, 21 June 2014</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the case of the E.CARU project, those characteristics where used with the purpose of remove sodium ions in order to desalinize water, but nhaS can be used in many different forms and in other aspects of biotechnology.</p></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><p>In the case of the E.CARU project, those characteristics where used with the purpose of remove sodium ions in order to desalinize water, but nhaS can be used in many different forms and in other aspects of biotechnology.</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>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div> </div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><br></ins></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><p><del class="diffchange diffchange-inline">2) </del>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor and if the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch actually worked at 35°C. In the case of the plaque that had the odor even if it was incubate at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of winter green enzyme by saturating it. </p></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline"><p><b>Aroma module</b><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><p>In the Aroma module tests, the medium with 10mM of salicylic acid was the one that produced the most intense odor and if the quantity is increased to 30m the odor is not reported. This indicates that the plasmid was actually in the bacteria and that the riboswitch actually worked at 35°C. In the case of the plaque that had the odor even if it was incubate at 29 ºC, it is concluded that this happened because the riboswitch is very sensitive to heat, and it was activated during the little time at which it was outside the incubator. In the case of the bacterias exposed to salicylic acid at 30mM, they presented a very similar odor despite of other conditions. This means that the 30mM concentration of salicylic acid could eliminate bateria or could affect the enzymatic reaction of winter green enzyme by saturating it. </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>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="text-align: right;"><a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_conclusions#"><font color="blue">Return to the Top</font></a></p></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="text-align: right;"><a href="https://2014hs.igem.org/Team:CIDEB-UANL_Mexico/labwork_conclusions#"><font color="blue">Return to the Top</font></a></p></div></div></td></tr>
</table>Marea