http://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&feed=atom&action=historyTeam:Harvard/Laser - Revision history2024-03-28T17:26:17ZRevision history for this page on the wikiMediaWiki 1.16.5http://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=169013&oldid=prevAgoyal at 03:53, 22 October 20092009-10-22T03:53:31Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As is visible below, there were a number of colonies that demonstrated beta-gal production only in the presence of light and PCB. We concluded that these cell lines contained the desired functional PhyB/PIF3 two hybrid system. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As is visible below, there were a number of colonies that demonstrated beta-gal production only in the presence of light and PCB. We concluded that these cell lines contained the desired functional PhyB/PIF3 two hybrid system. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Screen assay—petri dishes</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Screen assay—petri dishes</b></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For colony lifts from solid agar, nitrocellulose filters were used. After cutting an appropriately sized filter to fit the surface of a petri plate, nitrocellulose was laid on the agar surface and then removed using tweezers. The filters were then placed onto an aluminum sheet that was floated on liquid nitrogen, for approx. 20 seconds. The filters were then immersed in liquid nitrogen for 2 seconds, thawed at room temperature. Filter lifts were then placed cell side up onto pads of Whatman #1 filter paper, in empty petri-plates, that had been soaked with Z-buffer solution containing 2-mercaptoethanol and X-gal.The filter lifts were then incubated at 30 C for 30min to overnight, until the development of a blue precipitate was clearly visible.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For colony lifts from solid agar, nitrocellulose filters were used. After cutting an appropriately sized filter to fit the surface of a petri plate, nitrocellulose was laid on the agar surface and then removed using tweezers. The filters were then placed onto an aluminum sheet that was floated on liquid nitrogen, for approx. 20 seconds. The filters were then immersed in liquid nitrogen for 2 seconds, thawed at room temperature. Filter lifts were then placed cell side up onto pads of Whatman #1 filter paper, in empty petri-plates, that had been soaked with Z-buffer solution containing 2-mercaptoethanol and X-gal.The filter lifts were then incubated at 30 C for 30min to overnight, until the development of a blue precipitate was clearly visible.</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>For the initial screen, after transformation with plasmids containing the PhyB-DBD and Pif3-AD, plates were pre-incubated overnight , in the dark, at 30 deg. C with 25 <del class="diffchange diffchange-inline">M </del>PCB or DMSO only. After replica plating onto similar media, the plates were then incubated overnight, under red light. Positive colonies were then picked that expressed beta-galactosidase only under red-light induction. These were then re-streaked onto the same leu- trp- selection media and rescreened under overnight growth in red light, in the presence of PCB.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>For the initial screen, after transformation with plasmids containing the PhyB-DBD and Pif3-AD, plates were pre-incubated overnight , in the dark, at 30 deg. C with 25 <ins class="diffchange diffchange-inline">uM </ins>PCB or DMSO only. After replica plating onto similar media, the plates were then incubated overnight, under red light. Positive colonies were then picked that expressed beta-galactosidase only under red-light induction. These were then re-streaked onto the same leu- trp- selection media and rescreened under overnight growth in red light, in the presence of PCB.</div></td></tr>
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</table>Agoyalhttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=168914&oldid=prevNeena at 03:50, 22 October 20092009-10-22T03:50:53Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Incubation Time After Light Induction</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Incubation Time After Light Induction</b></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the reporter expression time course analysis, cells were grown in liquid culture in the same conditions as mentioned for the PCB concentration experiment mentioned above. 150 uL of cells in liquid media were then transferred into individual wells in a 96 well microtiter dish. Cells were then pulsed for 10 seconds using a laser, incubated for either 30 minutes or 1 hour in the dark and then the individual wells of cells were transferred onto nitrocellulose filter paper using a “bio-dot” apparatus connected to a vacuum. Significantly higher expression levels of beta-galactosidase were detected after 1 hour of growth, post laser induction vs. 30 minutes.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the reporter expression time course analysis, cells were grown in liquid culture in the same conditions as mentioned for the PCB concentration experiment mentioned above. 150 uL of cells in liquid media were then transferred into individual wells in a 96 well microtiter dish. Cells were then pulsed for 10 seconds using a laser, incubated for either 30 minutes or 1 hour in the dark and then the individual wells of cells were transferred onto nitrocellulose filter paper using a “bio-dot” apparatus connected to a vacuum. Significantly higher expression levels of beta-galactosidase were detected after 1 hour of growth, post laser induction vs. 30 minutes.</div></td></tr>
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</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=168823&oldid=prevNeena at 03:48, 22 October 20092009-10-22T03:48:22Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Phycocyanobilin.</b> PhyB function requires the covalent attachment of the prosthetic group phycocyanobilin, or PCB. Normally, plants and algae produce this prosthetic group, phycocyanobilin, or PCB, which attaches to the PhyB auto-catalytically. However, since yeast cells don’t normally express Phytochrome B, they also do not have the enzymes required to create PCB. Thus PCB had to be added to cultures for PhyB to function. Because PCB is not commercially available, we had to extract PCB from Spirulina algae, obtained from the health food store The Vitamin Shoppe. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Phycocyanobilin.</b> PhyB function requires the covalent attachment of the prosthetic group phycocyanobilin, or PCB. Normally, plants and algae produce this prosthetic group, phycocyanobilin, or PCB, which attaches to the PhyB auto-catalytically. However, since yeast cells don’t normally express Phytochrome B, they also do not have the enzymes required to create PCB. Thus PCB had to be added to cultures for PhyB to function. Because PCB is not commercially available, we had to extract PCB from Spirulina algae, obtained from the health food store The Vitamin Shoppe. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the laser pulse duration assay, yeast cells were grown and plated to the same density as mentioned for the PCB concentration assay. Cells were then treated to several pulse durations of light using a ~650nm wavelength, <3mW laser. The pulse durations used were ~0.5 seconds, 10 seconds and 30 seconds. All laser pulse durations were successful in inducing reporter expression, with the highest induction occurring at 10 seconds.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the laser pulse duration assay, yeast cells were grown and plated to the same density as mentioned for the PCB concentration assay. Cells were then treated to several pulse durations of light using a ~650nm wavelength, <3mW laser. The pulse durations used were ~0.5 seconds, 10 seconds and 30 seconds. All laser pulse durations were successful in inducing reporter expression, with the highest induction occurring at 10 seconds.</div></td></tr>
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</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=168605&oldid=prevNeena at 03:43, 22 October 20092009-10-22T03:43:56Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A similar approach was used in the semi-quantitative assays. Alternatively, yeast cells from liquid culture were immobilized onto nitrocellulose filter sheets using a Bio-Rad “Bio-Dot” apparatus connected to a vacuum manifold. The immobilized cells were lysed using liquid nitrogen and incubated with x-gal containing buffer as outlined above. For the analysis of beta-galactosidase expression in cells grown in 96 well micro-titer dishes, cotton applicators were used to remove cells from each well and then immersed directly into liquid nitrogen. After thawing at room temperature, they were then incubated for ~30-overnight in wells containing 150 uL of X-gal buffer.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>A similar approach was used in the semi-quantitative assays. Alternatively, yeast cells from liquid culture were immobilized onto nitrocellulose filter sheets using a Bio-Rad “Bio-Dot” apparatus connected to a vacuum manifold. The immobilized cells were lysed using liquid nitrogen and incubated with x-gal containing buffer as outlined above. For the analysis of beta-galactosidase expression in cells grown in 96 well micro-titer dishes, cotton applicators were used to remove cells from each well and then immersed directly into liquid nitrogen. After thawing at room temperature, they were then incubated for ~30-overnight in wells containing 150 uL of X-gal buffer.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>PCB Concentration Assay</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>PCB Concentration Assay</b></div></td></tr>
</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=168096&oldid=prevNeena at 03:32, 22 October 20092009-10-22T03:32:20Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2009.igem.org/Team:Harvard/PCB">PCB Biosynthesis in Yeast</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2009.igem.org/Team:Harvard/PCB">PCB Biosynthesis in Yeast</a></li></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2009.igem.org/Team:Harvard/Split">Split Luciferase</a></li></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <li><a href="https://2009.igem.org/Team:Harvard/Split">Split Luciferase</a></li></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/Parts">Parts</a></p></h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/Parts">Parts</a></p></h2></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/References">References</a></p></h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/References">References</a></p></h2></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/Ethics">Ethics</a></h2></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> <h2><p><a href="https://2009.igem.org/Team:Harvard/Ethics">Ethics</a></h2></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;"> <h2><p><a href="https://2009.igem.org/Team:Harvard/Safety">Safety</a></h2></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;"> <h2><p><a href="https://2009.igem.org/Team:Harvard/Acknowledgements">Acknowledgements</a></h2></ins></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </td></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div> </td></div></td></tr>
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</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=167876&oldid=prevNeena at 03:23, 22 October 20092009-10-22T03:23:23Z<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;"><br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br> <br></br></div></td></tr>
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</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=167767&oldid=prevNeena at 03:19, 22 October 20092009-10-22T03:19:19Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>PCB Concentration Assay</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>PCB Concentration Assay</b></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the PCB concentration assay, Y190 yeast cells containing the PhyB-DBD/Pif3-AD system were grown in selective leu- trp- liquid media to a density of 1 X106 cells/ml and 1 X 104 cells were plated in each well of a 96 well microtiter plate, containing 100 uL of solid media containing differing concentrations of PCB: 10 uM, 25 uM and 50 uM or equivalent volumes of DMSO carrier. All tested concentrations of PCB proved effective in inducing lacZ expression.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the PCB concentration assay, Y190 yeast cells containing the PhyB-DBD/Pif3-AD system were grown in selective leu- trp- liquid media to a density of 1 X106 cells/ml and 1 X 104 cells were plated in each well of a 96 well microtiter plate, containing 100 uL of solid media containing differing concentrations of PCB: 10 uM, 25 uM and 50 uM or equivalent volumes of DMSO carrier. All tested concentrations of PCB proved effective in inducing lacZ expression.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><<del class="diffchange diffchange-inline">b</del>><del class="diffchange diffchange-inline">***Insert Picture 96 well plate pcb concentration***</del></<del class="diffchange diffchange-inline">b</del>><br></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><<ins class="diffchange diffchange-inline">div class="center"><img src="https://static.igem.org/mediawiki/2009/c/ca/CORRECT_initial_light_induction_data.jpg" width="350" height="275"</ins>/><ins class="diffchange diffchange-inline">,</ins><<ins class="diffchange diffchange-inline">/div></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the laser pulse duration assay, yeast cells were grown and plated to the same density as mentioned for the PCB concentration assay. Cells were then treated to several pulse durations of light using a ~650nm wavelength, <3mW laser. The pulse durations used were ~0.5 seconds, 10 seconds and 30 seconds. All laser pulse durations were successful in inducing reporter expression, with the highest induction occurring at 10 seconds.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the laser pulse duration assay, yeast cells were grown and plated to the same density as mentioned for the PCB concentration assay. Cells were then treated to several pulse durations of light using a ~650nm wavelength, <3mW laser. The pulse durations used were ~0.5 seconds, 10 seconds and 30 seconds. All laser pulse durations were successful in inducing reporter expression, with the highest induction occurring at 10 seconds.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>***Insert q-tip laser pulse duration Assay***</b><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br><b>***Insert q-tip laser pulse duration Assay***</b><br></div></td></tr>
</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=167589&oldid=prevNeena at 03:14, 22 October 20092009-10-22T03:14:51Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As is visible below, there were a number of colonies that demonstrated beta-gal production only in the presence of light and PCB. We concluded that these cell lines contained the desired functional PhyB/PIF3 two hybrid system. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As is visible below, there were a number of colonies that demonstrated beta-gal production only in the presence of light and PCB. We concluded that these cell lines contained the desired functional PhyB/PIF3 two hybrid system. </div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del style="color: red; font-weight: bold; text-decoration: none;"><div class="center"><img src="https://static.igem.org/mediawiki/2009/9/98/Colony_light_screen.jpg" width="500" height="266"/>,</div></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><b>Screen assay—petri dishes</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Screen assay—petri dishes</b></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the initial screen, after transformation with plasmids containing the PhyB-DBD and Pif3-AD, plates were pre-incubated overnight , in the dark, at 30 deg. C with 25 M PCB or DMSO only. After replica plating onto similar media, the plates were then incubated overnight, under red light. Positive colonies were then picked that expressed beta-galactosidase only under red-light induction. These were then re-streaked onto the same leu- trp- selection media and rescreened under overnight growth in red light, in the presence of PCB.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For the initial screen, after transformation with plasmids containing the PhyB-DBD and Pif3-AD, plates were pre-incubated overnight , in the dark, at 30 deg. C with 25 M PCB or DMSO only. After replica plating onto similar media, the plates were then incubated overnight, under red light. Positive colonies were then picked that expressed beta-galactosidase only under red-light induction. These were then re-streaked onto the same leu- trp- selection media and rescreened under overnight growth in red light, in the presence of PCB.</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><br><<del class="diffchange diffchange-inline">b</del>><del class="diffchange diffchange-inline">***Insert Picture about yeast screen plates***</del></<del class="diffchange diffchange-inline">b</del>><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><<ins class="diffchange diffchange-inline">/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 class="diffchange diffchange-inline">div class="center"><img src="https://static.igem.org/mediawiki/2009/9/98/Colony_light_screen.jpg" width="500" height="266"</ins>/><ins class="diffchange diffchange-inline">,</ins><<ins class="diffchange diffchange-inline">/div></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 class="diffchange diffchange-inline"><br></</ins>br></div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><<del class="diffchange diffchange-inline">b</del>><del class="diffchange diffchange-inline">***Insert Picture about restreaked colonies***</del></<del class="diffchange diffchange-inline">b</del>><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><<ins class="diffchange diffchange-inline">/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 class="diffchange diffchange-inline">div class="center"><img src="https://static.igem.org/mediawiki/2009/e/e3/Picked_positive_colony_from_screen_plate.JPG" width="400" height="300"</ins>/><ins class="diffchange diffchange-inline">,</ins><<ins class="diffchange diffchange-inline">/div></ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><HR></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><HR></div></td></tr>
</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=167447&oldid=prevNeena at 03:11, 22 October 20092009-10-22T03:11:33Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><<del class="diffchange diffchange-inline">b</del>><del class="diffchange diffchange-inline">***Insert Picture Colony Screen***</del></<del class="diffchange diffchange-inline">b</del>><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><<ins class="diffchange diffchange-inline">/br</ins>></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Screen assay—petri dishes</b></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><b>Screen assay—petri dishes</b></div></td></tr>
</table>Neenahttp://2009.igem.org/wiki/index.php?title=Team:Harvard/Laser&diff=167376&oldid=prevNeena at 03:09, 22 October 20092009-10-22T03:09:02Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><br><<del class="diffchange diffchange-inline">b</del>><del class="diffchange diffchange-inline">***Insert Picture PCB Growth Curve***</del></<del class="diffchange diffchange-inline">b</del>><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div> </div></td></tr>
<tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><br><<ins class="diffchange diffchange-inline">/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 class="diffchange diffchange-inline">div class="center"><img src="https://static.igem.org/mediawiki/2009/3/3b/PCB_Growth_Curve.png" width="420" height="320"</ins>/><ins class="diffchange diffchange-inline">,</ins><<ins class="diffchange diffchange-inline">/div></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 class="diffchange diffchange-inline"><br></</ins>br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><br></div></td></tr>
</table>Neena