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</ol> | </ol> | ||
<ul> | <ul> | ||
− | <li> Instead of making up new competent cells, we used commercial competent cells which proved successful as we were able to grow HydEFG transformants. Plasmids from these were extracted via miniprep. They were digested and run on a gel. The gel results were not as expected _____. Because of this, new colonies should be screened. </li> | + | <li> Instead of making up new competent cells, we used commercial competent cells which proved successful as we were able to grow HydEFG transformants. </li> |
− | <li> The sequencing came back for Fer/Hyd A and B. Both were as confirmed. | + | <ul> <li> Plasmids from these were extracted via miniprep. </li> |
− | The glycerol stocks were then used to prep overnight cultures which were induced and tested for their ability to produce hydrogen. | + | <li> They were digested and run on a gel. </li> |
− | The Clark electrode was used to measure the H2 production and recordings went off the scale. This was great news. We are looking into the best ways to present this data. </li> | + | <li> The gel results were not as expected _____. Because of this, new colonies should be screened. </li> |
+ | </ul> | ||
+ | <li> The sequencing came back for Fer/Hyd A and B. Both were as confirmed. </li> | ||
+ | <li> The glycerol stocks were then used to prep overnight cultures which were induced and tested for their ability to produce hydrogen. </li> | ||
+ | <li> The Clark electrode was used to measure the H2 production and recordings went off the scale. This was great news. We are looking into the best ways to present this data. </li> | ||
<li> Unfortunately we still have not had any success with PCR amplification of backbones.</li> | <li> Unfortunately we still have not had any success with PCR amplification of backbones.</li> | ||
<li>HydEFG were sent to be sequenced and other cultures were grown as well as their glycerol stock. </li> | <li>HydEFG were sent to be sequenced and other cultures were grown as well as their glycerol stock. </li> | ||
</ul> | </ul> | ||
− | |||
</td> | </td> | ||
+ | </tr> | ||
</table> | </table> | ||
</p> | </p> | ||
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<td> <top> <img class="media object pull-left" src="https://static.igem.org/mediawiki/2017/c/c8/Notepadlogo.jpg"> </top> </td> | <td> <top> <img class="media object pull-left" src="https://static.igem.org/mediawiki/2017/c/c8/Notepadlogo.jpg"> </top> </td> | ||
<td> | <td> | ||
+ | <ul> | ||
+ | <li> | ||
+ | </li> | ||
</td> | </td> | ||
+ | </tr> | ||
<tr> | <tr> | ||
<td> <top> <img class="media object pull-left" src="https://static.igem.org/mediawiki/2017/8/81/T--Macquarie_Australia--flasklogo_.png" alt="flasklogo" style= "height:50%; width:50%"> | <td> <top> <img class="media object pull-left" src="https://static.igem.org/mediawiki/2017/8/81/T--Macquarie_Australia--flasklogo_.png" alt="flasklogo" style= "height:50%; width:50%"> | ||
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<li> </li> | <li> </li> | ||
</ul> | </ul> | ||
+ | </td> | ||
</tr> | </tr> | ||
− | |||
<tr> | <tr> | ||
<td> <top> <img src="https://static.igem.org/mediawiki/2017/8/81/T--Macquarie_Australia--flasklogo_.png" alt="flasklogo" style= "height:50%; width:50%"> | <td> <top> <img src="https://static.igem.org/mediawiki/2017/8/81/T--Macquarie_Australia--flasklogo_.png" alt="flasklogo" style= "height:50%; width:50%"> | ||
</td> | </td> | ||
<td> <h4> Wet Lab </h4> | <td> <h4> Wet Lab </h4> | ||
− | |||
<ul> | <ul> | ||
− | <li> </li> | + | <li> This week had a major focus on completing the Interlab study (Adrianna, Winonah, Jocelyn, Ali, Adrianna, Indi, Ed and Thi were involved)</li> |
+ | <ul> <li> Transformations and overnight cultures were completed in time for the big day of data collection on Thursday which included the Interlab fluorescence measurement of plates. </li> | ||
+ | <li> We started early on the interlab task, first we made fresh cell cultures from the cells that had been incubating over night. Then we started with the absorbance measurements of _____. We then made our time 0 samples, put the new cell cultures in the incubating shaker and left for 2 hours. </li> | ||
+ | <ul> <li> During the 2 hour period the serial dilution plate was set up ____ </li> </ul> | ||
+ | <li> Towards the end of interlab day, Team Member numbers were dwindling so Ed and Thi assisted with some pipetting to ensure we would be able to obtain spectrometer readings that day. </li> </ul> | ||
+ | <li> At the same time, a backbone swap to CAM for Fer/Hyd took place followed by transformation and plating up. </li> | ||
+ | <li> Hydrogen production data collection by Clark electrode continued. </li> | ||
+ | <li> HydEFG was digested (single and double digests performed on 7 separate colonies) and screened on gel (#V, 45 mins). Meanwhile we await sequencing result. </li> | ||
+ | <ul> <li> As the gel was running a team member messaged the group saying that unfortunately the protocol used was not an updated version… </li> | ||
+ | <li> After visualising the completed gel, it appeared the digests were only successful for 3/7 samples? </li> </ul> | ||
+ | <li> No more backbone PCR amplifications took place this week.</li> </ul> | ||
</ul> | </ul> | ||
+ | </td> | ||
+ | </tr> | ||
</table> | </table> | ||
</p> | </p> | ||
+ | |||
<h2> Week 8 (21/8/17-25/8/17) </h2> | <h2> Week 8 (21/8/17-25/8/17) </h2> | ||
<p> | <p> |
Revision as of 14:35, 10 September 2017
Notebook
Glossary
Week 1 (3/7/17-7/7/17)
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Dry Lab
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Week 2 (10/7/17-14/7/17)
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Dry Lab
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Wet Lab
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Week 3 (17/7/17-21/7/17)
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Dry Lab
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Wet Lab
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Week 4 (24/7/17-28/7/17)
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Dry Lab
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Wet Lab
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Week 5 (31/7/17-4/7/17)
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Dry Lab
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Wet Lab
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Week 6 (7/8/17-11/8/17)
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Dry Lab
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Wet LabOur plan was as follows:
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Week 7 (14/8/17-18/8/17)
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Dry Lab |
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Wet Lab
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Week 8 (21/8/17-25/8/17)
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Dry Lab |
Wet Lab |
Week 9 (28/8/17-1/9/17)
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Dry Lab |
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Wet Lab |
Week 10 (4/9/17-8/9/17)
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Dry Lab |
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Wet Lab |
Week 11 (22/9/17-15/9/17)
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Dry Lab |
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Wet Lab |
Week 12 (18/9/17-22/9/17)
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Dry Lab |
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Wet Lab |
Week 13 (25/9/17-29/9/17)
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Dry Lab |
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Wet Lab |
Week 14 (2/10/17-6/10/17)
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Dry Lab |
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Wet Lab
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Week 15 (9/10/17-13/10/17)
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Dry Lab> |
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Wet Lab |
Week 16 (16/10/17-20/10/17)
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Dry Lab |
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Wet Lab |
Week 17 (23/10/17-27/10/17)
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Dry Lab |
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Wet Lab |
Week 18 (30/10/17-3/10/17)
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Dry Lab |
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Wet Lab |