http://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&feed=atom&action=historyTeam:BCCS-Bristol/Wetlab/RFPbead tests - Revision history2024-03-28T16:08:17ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=192984&oldid=prevAmaty at 19:12, 27 October 20102010-10-27T19:12:05Z<p></p>
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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;">=Our Final Beads=</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:BCCS-Bristol/newtoc}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:BCCS-Bristol/newtoc}}</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;">==Motivation==</del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing ''E. coli'' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing ''E. coli'' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</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>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Spectro-photometer data==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Spectro-photometer data==</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:GFP RFP in beads.png|500px]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:GFP RFP in beads.png|500px]]</div></td></tr>
</table>Amatyhttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=191348&oldid=prevTtodd at 18:22, 27 October 20102010-10-27T18:22:54Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</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>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing 'E. coli' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing <ins class="diffchange diffchange-inline">'</ins>'E. coli<ins class="diffchange diffchange-inline">'</ins>' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Spectro-photometer data==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Spectro-photometer data==</div></td></tr>
</table>Ttoddhttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=190711&oldid=prevTtodd at 18:02, 27 October 20102010-10-27T18:02:39Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=Our Final Beads=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=Our Final Beads=</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</div></td></tr>
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</table>Ttoddhttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=185283&oldid=prevKcoyte at 15:01, 27 October 20102010-10-27T15:01:53Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>=<del class="diffchange diffchange-inline">RFP </del>Beads=</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>=<ins class="diffchange diffchange-inline">Our Final </ins>Beads=</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==Motivation==</div></td></tr>
</table>Kcoytehttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=144294&oldid=prevKcoyte at 13:23, 25 October 20102010-10-25T13:23:10Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=RFP Beads=</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>=RFP Beads=</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;">==Motivation==</ins></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>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing 'E. coli' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing 'E. coli' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.</div></td></tr>
</table>Kcoytehttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=136826&oldid=prevAmaty at 20:02, 24 October 20102010-10-24T20:02:47Z<p></p>
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</table>Amatyhttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=108246&oldid=prevKcoyte: /* RFP Beads */2010-10-18T11:10:35Z<p><span class="autocomment">RFP Beads</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:GFP RFP in beads.png|500px]]</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>[[Image:GFP RFP in beads.png|500px]]</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>
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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;">==Confocal Microscope Data==</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;">In an attempt to get better data on the behaviour of our ratio beads in soil, we then decided to examine them under a confocal microscope. This too indicated the presence of both GFP and RFP in the beads. </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;">Further, using a transect to average the fluorescence of the bacteria inside these beads indicated approximately twice as much GFP fluorescence in a bead taken from nitrogen saturated soil compared with non-nitrogenous soil. However this data is taken from only one bead, so should be treated cautiously.</ins></div></td></tr>
</table>Kcoytehttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=108234&oldid=prevKcoyte: /* Spectro-photometer data */2010-10-18T11:06:07Z<p><span class="autocomment">Spectro-photometer data</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[[Image:GFP RFP in beads.png|<del class="diffchange diffchange-inline">300px</del>]]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </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>[[Image:GFP RFP in beads.png|<ins class="diffchange diffchange-inline">500px</ins>]]</div></td></tr>
</table>Kcoytehttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=108233&oldid=prevKcoyte: /* Spectro-photometer data */2010-10-18T11:05:50Z<p><span class="autocomment">Spectro-photometer data</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>[Image:GFP RFP in beads.png|300px]</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[</ins>[Image:GFP RFP in beads.png|300px<ins class="diffchange diffchange-inline">]</ins>]</div></td></tr>
</table>Kcoytehttp://2010.igem.org/wiki/index.php?title=Team:BCCS-Bristol/Wetlab/RFPbead_tests&diff=108232&oldid=prevKcoyte: New page: =RFP Beads= Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced p...2010-10-18T11:05:20Z<p>New page: =RFP Beads= Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced p...</p>
<p><b>New page</b></p><div>=RFP Beads=<br />
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Having established our ratio method of signal detection worked well in soil, we next wanted to combine this with our new bead technology to form and test a yet more advanced prototype. We began by making beads containing 'E. coli' containing both BBa_K381001 and BBa_J04450 following the same method we had developed making GFP beads.<br />
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==Spectro-photometer data==<br />
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With beads made, we jumped straight to testing whether they worked on soil. The sets of beads were placed on non-sterile soil taken from a nearby field, some in petri-dishes with no added nitrogen, some on soil that had been saturated with 20mM of Potassium Nitrate. These beads were then left for 3 days to allow a signal to develop.<br />
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Unfortunately, unlike our previous experiments, we were not able to analyse signal using the stereo-microscope as an appropriate RFP filter was not available. Instead we were limited to collecting data using the photo-spectrometer. his proved extremely difficult due to the size and opacity of the beads, however, we were able to at least collect some data indicating that both GFP and RFP were expressed in the beads.<br />
<br />
The arrows in the graph below highlight the two peaks in absorbency indicating the presence of both GFP and RFP. Ideally we would like to get more quantitative data using a photo-spectrometer with relevant filters, however this was not possible within our timeframe.<br />
[Image:GFP RFP in beads.png|300px]</div>Kcoyte