http://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&feed=atom&action=historyTeam:Oxford/InterlabMeasurement - Revision history2024-03-29T00:24:55ZRevision history for this page on the wikiMediaWiki 1.16.5http://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=344766&oldid=prevCorinnaO: proofread2014-10-17T19:24:13Z<p>proofread</p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><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: #ffa; color:black; font-size: smaller;"><div><b>Single<del class="diffchange diffchange-inline">-</del>readings</b><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Single readings</b><br></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As before, 12 cultures were set up overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at 37°C until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As before, 12 cultures were set up overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at 37°C until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td></tr>
</table>CorinnaOhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=344743&oldid=prevCorinnaO at 19:23, 17 October 20142014-10-17T19:23:55Z<p></p>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><b>Time-course Protocol</b></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Time-course Protocol</b<ins class="diffchange diffchange-inline">><br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To understand the growth of the transformed DH5-a, we measured the Optical Density (OD), and corresponding Fluorescence (Fluo) for each device over the course of 40 cycles. This series of readings took over 13 hours, fully capturing the entire growth curve of the bacteria.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>To understand the growth of the transformed DH5-a, we measured the Optical Density (OD), and corresponding Fluorescence (Fluo) for each device over the course of 40 cycles. This series of readings took over 13 hours, fully capturing the entire growth curve of the bacteria.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3 cultures for each device were set-up overnight, taken from separate single colonies on the agar plate (biological replicates), as well as 3 cultures of non-transformed DH5-a (negative control). Of these 12 cultures, 3 repeated samples were taken to be read in our laboratory’s plate-reader. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3 cultures for each device were set-up overnight, taken from separate single colonies on the agar plate (biological replicates), as well as 3 cultures of non-transformed DH5-a (negative control). Of these 12 cultures, 3 repeated samples were taken to be read in our laboratory’s plate-reader. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We used an Infinite m2000 plate reader, with a Greiner 96 Flatback plate across all protocol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We used an Infinite m2000 plate reader, with a Greiner 96 Flatback plate across all protocol.</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <del class="diffchange diffchange-inline"><br></del><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><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: #ffa; color:black; font-size: smaller;"><div><b>Single-readings</b></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Single-readings</b<ins class="diffchange diffchange-inline">><br</ins>></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As before, 12 cultures were set up overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at 37°C until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>As before, 12 cultures were set up overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at 37°C until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td></tr>
</table>CorinnaOhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=344708&oldid=prevCorinnaO: proofread2014-10-17T19:23:28Z<p>proofread</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: #ffa; color:black; font-size: smaller;"><div><b>Time-course Protocol</b<del class="diffchange diffchange-inline">><br</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Time-course Protocol</b></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>To understand the growth of the transformed DH5-a, we measured the Optical Density (OD), and corresponding Fluorescence (Fluo) for each device over the course of 40 cycles. This series of readings took over 13 hours, fully capturing the <del class="diffchange diffchange-inline">bacteria’s </del>entire growth curve.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>To understand the growth of the transformed DH5-a, we measured the Optical Density (OD), and corresponding Fluorescence (Fluo) for each device over the course of 40 cycles. This series of readings took over 13 hours, fully capturing the entire growth curve <ins class="diffchange diffchange-inline">of the bacteria</ins>.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3 cultures for each device were set-up overnight, taken from separate single colonies on the agar plate (biological replicates), as well as 3 cultures of non-transformed DH5-a (negative control). Of these 12 cultures, 3 repeated samples were taken to be read in our laboratory’s plate-reader. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>3 cultures for each device were set-up overnight, taken from separate single colonies on the agar plate (biological replicates), as well as 3 cultures of non-transformed DH5-a (negative control). Of these 12 cultures, 3 repeated samples were taken to be read in our laboratory’s plate-reader. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We used an Infinite m2000 plate reader, with a Greiner 96 Flatback plate across all protocol.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We used an Infinite m2000 plate reader, with a Greiner 96 Flatback plate across all protocol.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><br><br></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><br><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: #ffa; color:black; font-size: smaller;"><div><b>Single-readings</b<del class="diffchange diffchange-inline">><br</del>></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><b>Single-readings</b></div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>As before, 12 cultures were <del class="diffchange diffchange-inline">setup </del>overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at <del class="diffchange diffchange-inline">37C </del>until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>As before, 12 cultures were <ins class="diffchange diffchange-inline">set up </ins>overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at <ins class="diffchange diffchange-inline">37°C </ins>until their OD reached, or was close to, 0.6. This took approximately 3 hours.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The amount of minimal media used for each culture was different dependent on the respective ODs. This was to standardise the amount of cells being used in the plate-reader. The specific volume of M9 salts was determined by (OD/0.6)ml. </div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>The amount of minimal media used for each culture was different dependent on the respective ODs. This was to standardise the amount of cells being used in the plate-reader. The specific volume of M9 salts was determined by (OD/0.6)ml. </div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Excitation 485nm, Emission 510nm</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Excitation 485nm, Emission 510nm</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Shaking before – 30 seconds</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Shaking before – 30 seconds</div></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Gain 2000<del class="diffchange diffchange-inline"><br></del><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Gain 2000<br><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: #ffa; color:black; font-size: smaller;"><div>This protocol was repeated the following day, using new individual colonies from the original agar plates. <del class="diffchange diffchange-inline"><br></del><br><br></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>This protocol was repeated the following day, using new individual colonies from the original agar plates. <br><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>We also took epifluorescent images of each cultures after the plate-readings, to visually inspect the variation in fluorescence per cell, and confirm that what we saw matched the data we were given by the plate-reader. For this we used a Nikon TE-200 microscope with GFP Filter set (Chroma), Hamamatsi EMCCD Camera and simple PCI software.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>We also took epifluorescent images of each cultures after the plate-readings, to visually inspect the variation in fluorescence per cell, and confirm that what we saw matched the data we were given by the plate-reader. For this we used a Nikon TE-200 microscope with GFP Filter set (Chroma), Hamamatsi EMCCD Camera and simple PCI software.</div></td></tr>
</table>CorinnaOhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=334673&oldid=prevJHoffman at 17:12, 17 October 20142014-10-17T17:12:24Z<p></p>
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</table>JHoffmanhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=329597&oldid=prevJHoffman at 15:53, 17 October 20142014-10-17T15:53:53Z<p></p>
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</table>JHoffmanhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=212783&oldid=prevTimAng94 at 11:18, 13 October 20142014-10-13T11:18:01Z<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">agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy agsdhg dhgsdkf sdfaskjdf hkjasdfhua duf asudoas dou saod sud vouas vouasod os oas oas dos au uf sfus fus fuyis fi sif yf iy</del></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><ins class="diffchange diffchange-inline"><b>Time-course Protocol</b><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">To understand the growth of the transformed DH5-a, we measured the Optical Density (OD), and corresponding Fluorescence (Fluo) for each device over the course of 40 cycles. This series of readings took over 13 hours, fully capturing the bacteria’s entire growth curve.</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">3 cultures for each device were set-up overnight, taken from separate single colonies on the agar plate (biological replicates), as well as 3 cultures of non-transformed DH5-a (negative control). Of these 12 cultures, 3 repeated samples were taken to be read in our laboratory’s plate-reader. </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">We used an Infinite m2000 plate reader, with a Greiner 96 Flatback plate across all protocol.</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">The resulting growth curves showed no peculiarities in either measurement, leading us to then decide to take single-readings of each culture at, or close to, the end of the log-phase. <br><br><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 class="diffchange diffchange-inline"><b>Single-readings</b><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">As before, 12 cultures were setup overnight. In the morning, 10ul were transferred from the overnight cultures into 5ml of LB broth, with appropriate amounts of antibiotic dependent on the device being used. These cultures were incubated and shaken at 37C until their OD reached, or was close to, 0.6. This took approximately 3 hours.</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">After this time, 1ml of each culture was centrifuged for 2 minutes, pipetting out all the supernatant. To re-suspend the pellet, we used 1X M9 salts so as to minimise autofluorescence contributing to our final readings. </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">The amount of minimal media used for each culture was different dependent on the respective ODs. This was to standardise the amount of cells being used in the plate-reader. The specific volume of M9 salts was determined by (OD/0.6)ml. </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">For each culture, 200ul were pipetted into the Greiner plate. This was repeated twice, giving two technical replicates and 24 wells occupied with samples. In addition, two wells were filled with M9 salts, as negative controls. Leaving the cover off, the plate was read once using the following settings:</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">Excitation 485nm, Emission 510nm</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">Shaking before – 30 seconds</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">Gain 2000<br><br><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 class="diffchange diffchange-inline">This protocol was repeated the following day, using new individual colonies from the original agar plates. <br><br><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 class="diffchange diffchange-inline">We also took epifluorescent images of each cultures after the plate-readings, to visually inspect the variation in fluorescence per cell, and confirm that what we saw matched the data we were given by the plate-reader. For this we used a Nikon TE-200 microscope with GFP Filter set (Chroma), Hamamatsi EMCCD Camera and simple PCI software.</ins></div></td></tr>
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</table>TimAng94http://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=198474&oldid=prevJHoffman: moved Team:Oxford/measurement to Team:Oxford/InterlabMeasurement2014-10-12T07:41:59Z<p>moved <a href="/Team:Oxford/measurement" class="mw-redirect" title="Team:Oxford/measurement">Team:Oxford/measurement</a> to <a href="/Team:Oxford/InterlabMeasurement" title="Team:Oxford/InterlabMeasurement">Team:Oxford/InterlabMeasurement</a></p>
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</table>JHoffmanhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=196642&oldid=prevJHoffman at 22:46, 11 October 20142014-10-11T22:46:11Z<p></p>
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</table>JHoffmanhttp://2014.igem.org/wiki/index.php?title=Team:Oxford/InterlabMeasurement&diff=196631&oldid=prevJHoffman at 22:45, 11 October 20142014-10-11T22:45:07Z<p></p>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2014/2/2b/Oxigemred.jpg" style="position:absolute; width:100%;z-index:-1; border-radius:15px;"/></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><img src="https://static.igem.org/mediawiki/2014/2/2b/Oxigemred.jpg" style="position:absolute; width:100%;z-index:-1; border-radius:15px;"/></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><div style="background-color:#D9D9D9; opacity:0.7; z-index:5<del class="diffchange diffchange-inline">; margin-left:7%</del>; Height:75px; width:<del class="diffchange diffchange-inline">500px</del>;font-size:65px;font-family:Helvetica;padding-top:5px; font-weight: 450;"></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><div style="background-color:#D9D9D9; opacity:0.7; z-index:5; Height:75px; width:<ins class="diffchange diffchange-inline">100%</ins>;font-size:65px;font-family:Helvetica;padding-top:5px; font-weight: 450<ins class="diffchange diffchange-inline">;margin-top:10px</ins>;"></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="background-color:white; opacity:0.7; Height:75px; width:100%;margin-top:5px:margin-bottom:5px;font-size:65px;font-family:Helvetica;padding-top:5px; color:#00000; font-weight: 450;"><br><font style="opacity:0.7">Measurement</font></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div><div style="background-color:white; opacity:0.7; Height:75px; width:100%;margin-top:5px:margin-bottom:5px;font-size:65px;font-family:Helvetica;padding-top:5px; color:#00000; font-weight: 450;"><br><font style="opacity:0.7">Measurement</font></div></div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></div></div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div></div></div></td></tr>
</table>JHoffman