http://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&feed=atom&action=historyTeam:RAMNOTIREN CALGARY/Project/Content - Revision history2024-03-29T10:49:10ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=26254&oldid=prevAhLicks at 01:43, 21 June 20142014-06-21T01:43:38Z<p></p>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=26206&oldid=prevAhLicks at 01:39, 21 June 20142014-06-21T01:39:44Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Muscles secrete lactic acid- the solution to this was for Central Memorials team to create a proof of concept idea. They would prove that bacteria with an anti Angiogenic factor such as angiostatin could be used to treat cancer cells. However in the future the sequence created could be inserted into a virus. This virus would contain a kill switch so that it would only be active and secreting angiostatin in the presence of a certain antibiotic. That way it could be used in conjunction with other treatment options. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Muscles secrete lactic acid- the solution to this was for Central Memorials team to create a proof of concept idea. They would prove that bacteria with an anti Angiogenic factor such as angiostatin could be used to treat cancer cells. However in the future the sequence created could be inserted into a virus. This virus would contain a kill switch so that it would only be active and secreting angiostatin in the presence of a certain antibiotic. That way it could be used in conjunction with other treatment options. <br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Bacteria in the body would create a sever immune response in an already sick body- A possible solution to this problem could be to insert the bacteria into the Nano tubes NASA is creating. These tubes will allow the angiostatin to be secreted out via the secretion tag, however the Nano capsule will contain the bacteria and keep the body from having an immune response to it.<br> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Bacteria in the body would create a sever immune response in an already sick body- A possible solution to this problem could be to insert the bacteria into the Nano tubes NASA is creating. These tubes will allow the angiostatin to be secreted out via the secretion tag, however the Nano capsule will contain the bacteria and keep the body from having an immune response to it.<br> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>• When the bacteria are injected into the body, how will it remain localized around the tumor? -This problem lead the team to an article explaining how a team of scientist at the University of California Berkeley, University of California and San Francisco have set out to create a tumor killing bacteria. This bacteria when injected into the bloodstream, would travel to the site of a tumor and insert itself into the cancer cell. Once inside the tumor it would produce a cancer- killing compound. In theory this cancer-killing compound could be our sequence. To read more about this bacteria click on this <a href="http://www.technologyreview.com/news/405899/tumor-killing-bacteria/">link</a><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>• When the bacteria are injected into the body, how will it remain localized around the tumor? -This problem lead the team to an article explaining how a team of scientist at the University of California Berkeley, University of California and San Francisco have set out to create a tumor killing bacteria. This bacteria when injected into the bloodstream, would travel to the site of a tumor and insert itself into the cancer cell. Once inside the tumor it would produce a cancer- killing compound. In theory this cancer-killing compound could be our sequence. To read more about this bacteria click on this <a href="http://www.technologyreview.com/news/405899/tumor-killing-bacteria/">link<ins class="diffchange diffchange-inline">.</ins></a><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Assays are designed and protocols carried out- Centrals team designed two assays that would be carried out the fallowing year to prove that their sequence would work. The first being one would show that the lactic acid promoter is active in the presence of lactic acid. Creating a circuit containing lactic acid and RFP would do this. To see exactly how this would be done proceed to the Assay section. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Assays are designed and protocols carried out- Centrals team designed two assays that would be carried out the fallowing year to prove that their sequence would work. The first being one would show that the lactic acid promoter is active in the presence of lactic acid. Creating a circuit containing lactic acid and RFP would do this. To see exactly how this would be done proceed to the Assay section. <br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Assays</h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Assays</h3></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>In different concentrations of lactic acid different amounts of red fluorescent protein should be produced.<br>This kit promotes angiogenesis or the formation of blood vessels. When this factor is active. circles are formed on the kit plate; after adding angiostatin to the kit plates, we hope the circles will disappear. This would show us the success of the angiostatin. In order to ensure that it is in fact our protein causing the circles to disappear, we will perform a few different tests involving the In Vitro Angiogenesis kit plate. First we will use the kit plate and add bacteria that do not contain our circuit. Hopefully the circles in the kit will stay intact, if they do, it will confirm that it is not our bacteria causing the circles to disappear. Next we will add bacteria containing our angiostatin that have not been lysed. If the cirlces remain in the plate, it confirms that our factor isn't leaking out of the bacteria. Finally, we will add our lysed bacteria to the kit plate (we will lyse the cells by adding detergent) and see if the circles disappear, if they do then our transformation of angiostatin into the bacteria was successful.<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>In different concentrations of lactic acid different amounts of red fluorescent protein should be produced.<br>This kit promotes angiogenesis or the formation of blood vessels. When this factor is active. circles are formed on the kit plate; after adding angiostatin to the kit plates, we hope the circles will disappear. This would show us the success of the angiostatin. In order to ensure that it is in fact our protein causing the circles to disappear, we will perform a few different tests involving the In Vitro Angiogenesis kit plate. First we will use the kit plate and add bacteria that do not contain our circuit. Hopefully the circles in the kit will stay intact, if they do, it will confirm that it is not our bacteria causing the circles to disappear. Next we will add bacteria containing our angiostatin that have not been lysed. If the cirlces remain in the plate, it confirms that our factor isn't leaking out of the bacteria. Finally, we will add our lysed bacteria to the kit plate (we will lyse the cells by adding detergent) and see if the circles disappear, if they do then our transformation of angiostatin into the bacteria was successful.<br></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The final essay performed will be performed in order to make sure that our circuit works correctly is quite simple. We will again use the kit plate we purchased that has an Angiogenic factor and put it into a well along with a concentration of lactic acid. This will be repeated five times with five different concentrations of lactic acid. If all goes well the circles should remain. In the next trial, everything will be kept the same; however, this time we will add bacteria. The next well will contain bacteria with our circuit inside. The circut, however,has not yet been lysed and therefore it will not be active. Again the kit plate and lactic acid is added to the well and it is repeated five times. In the last well, we will add our lysed bacteria and follow the same step we took for the previous sections. The well with the lowest concentration of lactic acid should have the greatest amount of circles however the amount of circles should be reduced compared to the wells where our circuit was not active. As the concentration of lactic acid increased the number of circles should <del class="diffchange diffchange-inline">dec</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The final essay performed will be performed in order to make sure that our circuit works correctly is quite simple. We will again use the kit plate we purchased that has an Angiogenic factor and put it into a well along with a concentration of lactic acid. This will be repeated five times with five different concentrations of lactic acid. If all goes well the circles should remain. In the next trial, everything will be kept the same; however, this time we will add bacteria. The next well will contain bacteria with our circuit inside. The circut, however,has not yet been lysed and therefore it will not be active. Again the kit plate and lactic acid is added to the well and it is repeated five times. In the last well, we will add our lysed bacteria and follow the same step we took for the previous sections. The well with the lowest concentration of lactic acid should have the greatest amount of circles however the amount of circles should be reduced compared to the wells where our circuit was not active. As the concentration of lactic acid increased the number of circles should <ins class="diffchange diffchange-inline">decrease</p></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><h3>What We Plan To Do</h3> </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>Our team plans to build three constructs. The first is a lactic acid promoter ligated with a red fluorescent protein. The second is a lactic acid promoter ligated with their angiostatin and the third is a lactic acid promoter ligated with angiostatin and a constituent promoter. </p><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins 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>In the first assay a circuit containing a constituent promoter will cause the angiostatin to be continuously expressed in the bacteria. We will then lyse it and then run them in a western. On the western, a dot or a line should appear which shows that angiostatin is being expressed in the bacteria.</p> <br></ins></div></td></tr>
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<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins style="color: red; font-weight: bold; text-decoration: none;"><p>The second assay is performed to test the lactic acid promoter. A circuit comprised of the lactic acid promoter ligated with red fluorescent protein is inserted into the bacteria. These bacteria are then grown in different concentrations of lactic acid and then lysed. Next they are run on a western, the brightest line on the western shows at which concentration the lactic acid has maximum expression.</p> <br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins 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>The third assay is executed to test the lactic acid promoter with the angiostatin. It is carried out to make sure that angiostatin is not toxic to the bacteria, or if it is toxic at what concentration is it toxic to the bacteria. The team also wants to see at what concentration of lactic acid could they get the maximum amount of angiostatin produced. To gather this information the team would put bacteria containing the ligated circuit containing a lactic acid promoter and angiostatin into different concentrations of lactic acid. The bacteria would be grown in these different concentrations of lactic acid and then lysed. Running a western would show Central Memorial at what concentration of lactic acid they got maximum induction of angiostatin (this would be the brightest line on the gel).</p><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins 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>The fourth assay carried out is done to show the concentration of lactic acid needed to induce enough angiostatin to inhibit angiogenesis. This is done by growing bacteria containing ligated circuits of lactic acid promoter to angiostatin in different concentrations of lactic acid. The bacteria are then lysed and added to a kit plates containing vascular growth cells. The plate with tubes formed out of these vascular growth cells show angiogenesis and the plates without tubes formed shows that growth of the vascular growth cells was inhibited. The well with the fewest tubes formed would show the concentration of angiostatin needed to inhibit angiogenesis and at what concentration of lactic acid the concentration of angiostatin was produced. </p><br></ins></div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins 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>The final assay performed would be carried to find a secretion tag capable of secreting the amount of angiostatin needed to inhibit angiogenesis. This would be done by putting a bacteria containing a circuit a ligated circuit of lactic acid promoter angiostatin and a secretion tag (several circuits would be created to put into bacteria containing different secretion tags) these bacteria would be grown in physiological concentrations of lactic acid (concentration of lactic acid produced by tumor cells), lysed and then run on a western. The secretion tag that secreted the concentration of angiostatin closest to that of the optimal amount needed to inhibit angiogenesis would choose. The optimal amount of angiostatin needed was determined in assay number four. The bacteria grown in the concentration of lactic acid producing optimal amounts of angiostatin (found in assay four) would be lysed on the far right corner of the western so that the secretion tag trials (which were run on the same western) could be compared to it.</p> <br></ins></div></td></tr>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25505&oldid=prevAhLicks at 23:46, 20 June 20142014-06-20T23:46:13Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Muscles secrete lactic acid- the solution to this was for Central Memorials team to create a proof of concept idea. They would prove that bacteria with an anti Angiogenic factor such as angiostatin could be used to treat cancer cells. However in the future the sequence created could be inserted into a virus. This virus would contain a kill switch so that it would only be active and secreting angiostatin in the presence of a certain antibiotic. That way it could be used in conjunction with other treatment options. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Muscles secrete lactic acid- the solution to this was for Central Memorials team to create a proof of concept idea. They would prove that bacteria with an anti Angiogenic factor such as angiostatin could be used to treat cancer cells. However in the future the sequence created could be inserted into a virus. This virus would contain a kill switch so that it would only be active and secreting angiostatin in the presence of a certain antibiotic. That way it could be used in conjunction with other treatment options. <br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Bacteria in the body would create a sever immune response in an already sick body- A possible solution to this problem could be to insert the bacteria into the Nano tubes NASA is creating. These tubes will allow the angiostatin to be secreted out via the secretion tag, however the Nano capsule will contain the bacteria and keep the body from having an immune response to it.<br> </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>• Bacteria in the body would create a sever immune response in an already sick body- A possible solution to this problem could be to insert the bacteria into the Nano tubes NASA is creating. These tubes will allow the angiostatin to be secreted out via the secretion tag, however the Nano capsule will contain the bacteria and keep the body from having an immune response to it.<br> </div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>• When the bacteria are injected into the body, how will it remain localized around the tumor? -This problem lead the team to an article explaining how a team of scientist at the University of California Berkeley, University of California and San Francisco have set out to create a tumor killing bacteria. This bacteria when injected into the bloodstream, would travel to the site of a tumor and insert itself into the cancer cell. Once inside the tumor it would produce a cancer- killing compound. In theory this cancer-killing compound could be our sequence. To read more about this bacteria click on <del class="diffchange diffchange-inline">the hyper link </del>http://www.technologyreview.com/news/405899/tumor-killing-bacteria/<br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>• When the bacteria are injected into the body, how will it remain localized around the tumor? -This problem lead the team to an article explaining how a team of scientist at the University of California Berkeley, University of California and San Francisco have set out to create a tumor killing bacteria. This bacteria when injected into the bloodstream, would travel to the site of a tumor and insert itself into the cancer cell. Once inside the tumor it would produce a cancer- killing compound. In theory this cancer-killing compound could be our sequence. To read more about this bacteria click on <ins class="diffchange diffchange-inline">this <a href="</ins>http://www.technologyreview.com/news/405899/tumor-killing-bacteria/<ins class="diffchange diffchange-inline">">link</a></ins><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Assays are designed and protocols carried out- Centrals team designed two assays that would be carried out the fallowing year to prove that their sequence would work. The first being one would show that the lactic acid promoter is active in the presence of lactic acid. Creating a circuit containing lactic acid and RFP would do this. To see exactly how this would be done proceed to the Assay section. <br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>6. Assays are designed and protocols carried out- Centrals team designed two assays that would be carried out the fallowing year to prove that their sequence would work. The first being one would show that the lactic acid promoter is active in the presence of lactic acid. Creating a circuit containing lactic acid and RFP would do this. To see exactly how this would be done proceed to the Assay section. <br></div></td></tr>
</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25500&oldid=prevAhLicks at 23:45, 20 June 20142014-06-20T23:45:06Z<p></p>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25499&oldid=prevAhLicks at 23:44, 20 June 20142014-06-20T23:44:33Z<p></p>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25495&oldid=prevAhLicks at 23:43, 20 June 20142014-06-20T23:43:11Z<p></p>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25480&oldid=prevAhLicks at 23:39, 20 June 20142014-06-20T23:39:38Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Our Idea</h3></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><h3>Our Idea</h3></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>Forty percent of Canadians are expected to develop cancer in their life time of that forty percent twenty five percent are expected to die from their cancer. Imagine what could be accomplished if all of those people were focusing their attention in their futures instead of the now? This question is exactly what motivated the Central Memorials IGEM team to create a bacterium that could target and kill cancer cells. We decided to combine the developing cancer treatment option of <a href="http://en.wikipedia.org/wiki/Angiogenesis">anti angiogenesis</a> with <a href="http://en.wikipedia.org/wiki/Synthetic_biology">synthetic biology</a> to create a biological machine that would cut off the blood supply feeding cancer cells, effectively starving them. This method of killing cancer cells inspired us to title our project "Choking Cancer".</p><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>Forty percent of Canadians are expected to develop cancer in their life time of that forty percent twenty five percent are expected to die from their cancer. Imagine what could be accomplished if all of those people were focusing their attention in their futures instead of the now? This question is exactly what motivated the Central Memorials IGEM team to create a bacterium that could target and kill cancer cells. We decided to combine the developing cancer treatment option of <a href="http://en.wikipedia.org/wiki/Angiogenesis">anti angiogenesis</a> with <a href="http://en.wikipedia.org/wiki/Synthetic_biology">synthetic biology</a> to create a biological machine that would cut off the blood supply feeding cancer cells, effectively starving them. This method of killing cancer cells inspired us to title our project "Choking Cancer".</p><br></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>In different concentrations of lactic acid different amounts of red fluorescent protein should be produced.<br>This kit promotes angiogenesis or the formation of blood vessels. When this factor is active. circles are formed on the kit plate; after adding angiostatin to the kit plates, we hope the circles will disappear. This would show us the success of the angiostatin. In order to ensure that it is in fact our protein causing the circles to disappear, we will perform a few different tests involving the In Vitro Angiogenesis kit plate. First we will use the kit plate and add bacteria that do not contain our circuit. Hopefully the circles in the kit will stay intact, if they do, it will confirm that it is not our bacteria causing the circles to disappear. Next we will add bacteria containing our angiostatin that have not been lysed. If the cirlces remain in the plate, it confirms that our factor isn't leaking out of the bacteria. Finally, we will add our lysed bacteria to the kit plate (we will lyse the cells by adding detergent) and see if the circles disappear, if they do then our transformation of angiostatin into the bacteria was successful.<br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <p>In different concentrations of lactic acid different amounts of red fluorescent protein should be produced.<br>This kit promotes angiogenesis or the formation of blood vessels. When this factor is active. circles are formed on the kit plate; after adding angiostatin to the kit plates, we hope the circles will disappear. This would show us the success of the angiostatin. In order to ensure that it is in fact our protein causing the circles to disappear, we will perform a few different tests involving the In Vitro Angiogenesis kit plate. First we will use the kit plate and add bacteria that do not contain our circuit. Hopefully the circles in the kit will stay intact, if they do, it will confirm that it is not our bacteria causing the circles to disappear. Next we will add bacteria containing our angiostatin that have not been lysed. If the cirlces remain in the plate, it confirms that our factor isn't leaking out of the bacteria. Finally, we will add our lysed bacteria to the kit plate (we will lyse the cells by adding detergent) and see if the circles disappear, if they do then our transformation of angiostatin into the bacteria was successful.<br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The final essay performed will be performed in order to make sure that our circuit works correctly is quite simple. We will again use the kit plate we purchased that has an Angiogenic factor and put it into a well along with a concentration of lactic acid. This will be repeated five times with five different concentrations of lactic acid. If all goes well the circles should remain. In the next trial, everything will be kept the same; however, this time we will add bacteria. The next well will contain bacteria with our circuit inside. The circut, however,has not yet been lysed and therefore it will not be active. Again the kit plate and lactic acid is added to the well and it is repeated five times. In the last well, we will add our lysed bacteria and follow the same step we took for the previous sections. The well with the lowest concentration of lactic acid should have the greatest amount of circles however the amount of circles should be reduced compared to the wells where our circuit was not active. As the concentration of lactic acid increased the number of circles should dec</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The final essay performed will be performed in order to make sure that our circuit works correctly is quite simple. We will again use the kit plate we purchased that has an Angiogenic factor and put it into a well along with a concentration of lactic acid. This will be repeated five times with five different concentrations of lactic acid. If all goes well the circles should remain. In the next trial, everything will be kept the same; however, this time we will add bacteria. The next well will contain bacteria with our circuit inside. The circut, however,has not yet been lysed and therefore it will not be active. Again the kit plate and lactic acid is added to the well and it is repeated five times. In the last well, we will add our lysed bacteria and follow the same step we took for the previous sections. The well with the lowest concentration of lactic acid should have the greatest amount of circles however the amount of circles should be reduced compared to the wells where our circuit was not active. As the concentration of lactic acid increased the number of circles should dec</div></td></tr>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25475&oldid=prevAhLicks at 23:38, 20 June 20142014-06-20T23:38:49Z<p></p>
<a href="http://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=25475&oldid=21978">Show changes</a>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=21978&oldid=prevAhLicks at 06:27, 20 June 20142014-06-20T06:27:02Z<p></p>
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</table>AhLickshttp://2014hs.igem.org/wiki/index.php?title=Team:RAMNOTIREN_CALGARY/Project/Content&diff=21961&oldid=prevAhLicks at 06:21, 20 June 20142014-06-20T06:21:58Z<p></p>
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