http://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&feed=atom&action=historyTeam:TU-Eindhoven/LabJournal - Revision history2024-03-28T20:36:34ZRevision history for this page on the wikiMediaWiki 1.16.5http://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=330803&oldid=prevZc.felix at 09:06, 26 October 20132013-10-26T09:06:08Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=The Western blot clearly shows the bands which were marked on the SDS gel as being EGFP. We also no longer see the thick upper bands showing that these are in fact not EGFP. Futher the columns are given as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in higher length| id=WesternBlot }}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=The Western blot clearly shows the bands which were marked on the SDS gel as being EGFP. We also no longer see the thick upper bands showing that these are in fact not EGFP. Futher the columns are given as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in higher length<ins class="diffchange diffchange-inline">.</ins>| id=WesternBlot }}</div></td></tr>
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</table>Zc.felixhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329092&oldid=prevArdjanvanderLinden: /* Western Blot */2013-10-25T16:15:31Z<p><span class="autocomment">Western Blot</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=The Western blot clearly shows the bands which were marked on the SDS gel as being EGFP. We also no longer see the thick upper bands showing that these are in fact not EGFP. Futher the columns are given as follows:| id=WesternBlot }}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=The Western blot clearly shows the bands which were marked on the SDS gel as being EGFP. We also no longer see the thick upper bands showing that these are in fact not EGFP. Futher the columns are given as follows: <ins class="diffchange diffchange-inline">1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in higher length</ins>| id=WesternBlot }}</div></td></tr>
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</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329089&oldid=prevArdjanvanderLinden: /* Coomaaise Analysis */2013-10-25T16:14:44Z<p><span class="autocomment">Coomaaise Analysis</span></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in <del class="diffchange diffchange-inline">hoigher </del>length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in <ins class="diffchange diffchange-inline">higher </ins>length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td></tr>
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</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329087&oldid=prevArdjanvanderLinden: /* Western Blot */2013-10-25T16:14:08Z<p><span class="autocomment">Western Blot</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>An image of the final western blot can be seen below:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>An image of the final western blot can be seen below:</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">{{:Team:TU-Eindhoven/Template:FloatEnd | caption=The Western blot clearly shows the bands which were marked on the SDS gel as being EGFP. We also no longer see the thick upper bands showing that these are in fact not EGFP. Futher the columns are given as follows:| id=WesternBlot }}</ins></div></td></tr>
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</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329082&oldid=prevArdjanvanderLinden: /* Fluorescence Measurements */2013-10-25T16:11:51Z<p><span class="autocomment">Fluorescence Measurements</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>From the purified proteins we now pippetted 200&micro;L of each sample into a well of a 96 wells plate. This plate was then left open so that the samples could mature and was subsequently placed in the fluorescence reader where the measurements were conducted. Using the same preferences as we had used on October 22nd we would be able to accurately compare the results of both EGFP expression. The results of the fluorescence measurements are shown below:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>From the purified proteins we now pippetted 200&micro;L of each sample into a well of a 96 wells plate. This plate was then left open so that the samples could mature and was subsequently placed in the fluorescence reader where the measurements were conducted. Using the same preferences as we had used on October 22nd we would be able to accurately compare the results of both EGFP expression. The results of the fluorescence measurements are shown below:</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=EGFP intensity over time| id=EGFP intensities over time }}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=EGFP intensity over time| id=EGFP intensities over time }}</div></td></tr>
</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329079&oldid=prevArdjanvanderLinden: /* Coomaaise Analysis */2013-10-25T16:11:09Z<p><span class="autocomment">Coomaaise Analysis</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Hereafter the gel was washed with de-mineralissed water for a number of hours before the plate was imaged:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Hereafter the gel was washed with de-mineralissed water for a number of hours before the plate was imaged:</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:Image | filename=AnaerobicEGFPExpressionPurifiedSDS.jpg}}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:Image | filename=AnaerobicEGFPExpressionPurifiedSDS.jpg}}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in hoigher length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in hoigher length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td></tr>
</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329078&oldid=prevArdjanvanderLinden: /* Coomaaise Analysis */2013-10-25T16:10:49Z<p><span class="autocomment">Coomaaise Analysis</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in hoigher length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in hoigher length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</div></td></tr>
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</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329075&oldid=prevArdjanvanderLinden: /* Coomaaise Analysis */2013-10-25T16:07:59Z<p><span class="autocomment">Coomaaise Analysis</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After having run and washed the gels in water one of the gels was placed in 1x used coomaaise medium. The gel in solution was then placed on a rocking table and left to stain for approximately 2 hours. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After having run and washed the gels in water one of the gels was placed in 1x used coomaaise medium. The gel in solution was then placed on a rocking table and left to stain for approximately 2 hours. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Hereafter the gel was washed with de-mineralissed water for a number of hours before the plate was imaged:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Hereafter the gel was washed with de-mineralissed water for a number of hours before the plate was imaged:</div></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">{{:Team:TU-Eindhoven/Template:ImageList}}</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">{{:Team:TU-Eindhoven/Template:Float | position=left | size=6 }}</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">{{:Team:TU-Eindhoven/Template:Image | filename=AnaerobicEGFPExpressionPurifiedSDS}}</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">{{:Team:TU-Eindhoven/Template:FloatEnd | caption=SDS gel showing the results of loading the purified EGFP samples onto a 12% SDS gel. The columns contain as follows: 1: Aerobic Control, 2-7: Samples taken during the anaerobic expression at 0% Oxygen saturation, 8-13: Samples taken during the anaerobic expression at 5% Oxygen saturation, 14: Control EGFP bound to CNA35 resulting in hoigher length (Control for Westernblot). The EGFP protein bands of interest are shown in red, the higher thicker bands above do not represent EGFP as later shown by the western blot, futher more the size indicated by these bands does not fit the EGFP profile.| id=1G70_Double_Bands }}</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Western Blot====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Western Blot====</div></td></tr>
</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329071&oldid=prevArdjanvanderLinden: /* Fluorescence Measurements */2013-10-25T16:02:54Z<p><span class="autocomment">Fluorescence Measurements</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=EGFP intensity over time| id=EGFP intensities over time }}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{{:Team:TU-Eindhoven/Template:FloatEnd | caption=EGFP intensity over time| id=EGFP intensities over time }}</div></td></tr>
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</table>ArdjanvanderLindenhttp://2013.igem.org/wiki/index.php?title=Team:TU-Eindhoven/LabJournal&diff=329056&oldid=prevArdjanvanderLinden: /* Fluorescence Measurements */2013-10-25T15:59:33Z<p><span class="autocomment">Fluorescence Measurements</span></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Fluorescence Measurements====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====Fluorescence Measurements====</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>From the purified proteins we now pippetted 200&micro;L of each sample into a well of a 96 wells plate. This plate was then left open so that the samples could mature and was subsequently placed in the fluorescence reader where the measurements were conducted. Using the same preferences as we had used on October 22nd we would be able to accurately compare the results of both EGFP expression. The results of the fluorescence measurements are shown below:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>From the purified proteins we now pippetted 200&micro;L of each sample into a well of a 96 wells plate. This plate was then left open so that the samples could mature and was subsequently placed in the fluorescence reader where the measurements were conducted. Using the same preferences as we had used on October 22nd we would be able to accurately compare the results of both EGFP expression. The results of the fluorescence measurements are shown below:</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;">{{:Team:TU-Eindhoven/Template:FloatEnd | caption=EGFP intensity over time| id=EGFP intensities over time }}</ins></div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====SDS Gel and Western Blot Preparation====</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>====SDS Gel and Western Blot Preparation====</div></td></tr>
</table>ArdjanvanderLinden