Difference between revisions of "Team:Reading/Notebook"

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<p> <b> 07/07/15 </b> </p>
 
<p> <b> 07/07/15 </b> </p>
 
<li>Flask 5 cultured containing 10ul <i>6803</i> WT in 100ml BG-11. This flask will be used for preliminary growth curve measurements. The flask was placed on the shaker with flasks 2,3 and 4 all at 58rpm and 30°C under lab overhead lights. </li>
 
<li>Flask 5 cultured containing 10ul <i>6803</i> WT in 100ml BG-11. This flask will be used for preliminary growth curve measurements. The flask was placed on the shaker with flasks 2,3 and 4 all at 58rpm and 30°C under lab overhead lights. </li>
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<table style="width:50%">
 
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<tr> <th> Date </th> <th> OD<sub>750</sub> </th> <th> Estimated Cells per ml </th> </tr>
 
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<tr> <td> 07/07 </td> <td> 0.000 </td> <td> 0 </td> </tr>
 
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<tr> <td> 08/07 </td> <td> 0.003 </td> <td> 4.8x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 09/07 </td> <td> 0.005 </td> <td> 8x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 10/07 </td> <td> 0.005 </td> <td> 8x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 13/07 </td> <td> 0.005 </td> <td> 8x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 14/07 </td> <td> 0.112 </td> <td> 1.792x10<sup>7</sup> </td> </tr>
 
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<tr> <td> 15/07 </td> <td> 0.008 </td> <td> 1.28x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 16/07 </td> <td> 0.009 </td> <td> 1.44x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 17/07 </td> <td> 0.030 </td> <td> 4.8x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 20/07 </td> <td> 0.364 </td> <td> 5.824x10<sup>7</sup> </td> </tr>
 
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<tr> <td> 21/07 </td> <td> 0.510 </td> <td> 8.160x10<sup>7</sup> </td> </tr>
 
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<tr> <td> 22/07 </td> <td> 0.820 </td> <td> 1.312x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 23/07 </td> <td> 1.016 </td> <td> 1.626x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 24/07 </td> <td> 1.167 </td> <td> 1.876x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 27/07 </td> <td> 1.412 </td> <td> 2.259x10<sup>8</sup> </td> </tr>
 
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</table>
 
 
<p> <b> 10/07/15 </b> </p>  
 
<p> <b> 10/07/15 </b> </p>  
 
<p> 2 Litres of water collected from Whiteknights Lake on campus for SODIS, method outlines in protocols. </p>
 
<p> 2 Litres of water collected from Whiteknights Lake on campus for SODIS, method outlines in protocols. </p>
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<p> Estimating the Cells per ml for the flask from 29/06 1 a.u at 750nm = 1.6x10<sup>8</sup> <li> OD<sub>750</sub> = 0.028 </li> Therefore <li> 0.028x1.6x10<sup>8</sup> = 4.48x10<sup>6</sup> cells per ml </li> </p>  
 
<p> Estimating the Cells per ml for the flask from 29/06 1 a.u at 750nm = 1.6x10<sup>8</sup> <li> OD<sub>750</sub> = 0.028 </li> Therefore <li> 0.028x1.6x10<sup>8</sup> = 4.48x10<sup>6</sup> cells per ml </li> </p>  
 
<p> OD<sub>750</sub> measurement and cells per ml estimated for flasks cultured on 02/07 </p>
 
<p> OD<sub>750</sub> measurement and cells per ml estimated for flasks cultured on 02/07 </p>
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<table style="width:50%">
 
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<tr> <th> Date </th> <th> Flask No. </th> <th> OD<sub>750</sub> </th> <th> Estimated Cells per ml </th> </tr>
 
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<tr> <td rowspan="3"> 13/07 </td> <td> 2 </td> <td> 0.006 </td> <td> 9.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.012 </td> <td> 1.92x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.015 </td> <td> 2.4x10<sup>6</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 14/07 </td> <td> 2 </td> <td> 0.006 </td> <td> 9.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.018 </td> <td> 2.88x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.017 </td> <td> 2.72x10<sup>6</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 15/07 </td> <td> 2 </td> <td> 0.006 </td> <td> 9.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.021 </td> <td> 3.36x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.018 </td> <td> 2.88x10<sup>6</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 16/07 </td> <td> 2 </td> <td> 0.001 </td> <td> 1.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.020 </td> <td> 3.2x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.019 </td> <td> 3.04x10<sup>6</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 17/07 </td> <td> 2 </td> <td> 0.001 </td> <td> 1.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.040 </td> <td> 6.4x10<sup>6</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.027 </td> <td> 4.32x10<sup>6</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 20/07 </td> <td> 2 </td> <td> 0.005 </td> <td> 8x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.577 </td> <td> 9.232x10<sup>7</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.486 </td> <td> 7.776x10<sup>7</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 21/07 </td> <td> 2 </td> <td> 0.001 </td> <td> 1.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 0.757 </td> <td> 1.211x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.653 </td> <td> 1.045x10<sup>8</sup> </td> </tr>
 
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<tr> <td rowspan="3"> 22/07 </td> <td> 2 </td> <td> 0.001 </td> <td> 1.6x10<sup>5</sup> </td> </tr>
 
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<tr> <td> 3 </td> <td> 1.049 </td> <td> 1.678x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 0.916 </td> <td> 1.466x10<sup>8</sup> </td> </tr>
 
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<tr> <td rowspan="2"> 23/07 </td> <td> 3 </td> <td> 1.302 </td> <td> 2.083x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 1.203 </td> <td> 1.925x10<sup>8</sup> </td> </tr>
 
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<tr> <td rowspan="2"> 24/07 </td> <td> 3 </td> <td> 1.324 </td> <td> 2.118x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 1.296 </td> <td> 2.074x10<sup>8</sup> </td> </tr>
 
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<tr> <td rowspan="2"> 27/07 </td> <td> 3 </td> <td> 1.691 </td> <td> 2.706x10<sup>8</sup> </td> </tr>
 
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<tr> <td> 4 </td> <td> 1.650 </td> <td> 2.640x10<sup>8</sup> </td> </tr>
 
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</table>
 
 
<p> <b> 14/07/15 </b> </p>
 
<p> <b> 14/07/15 </b> </p>
 
<p> The lamp safety testing completed so flasks placed under the bright white light bulb </p>
 
<p> The lamp safety testing completed so flasks placed under the bright white light bulb </p>
 
<p> <b> 21/07/15 </b> </p>
 
<p> <b> 21/07/15 </b> </p>
 
<p> Flasks are getting very green, definitely a good sign! </p>  
 
<p> Flasks are getting very green, definitely a good sign! </p>  
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<p> <b> 23/07/15 </b> </p>
 
<p> <b> 23/07/15 </b> </p>
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<p> Flask 2 discarded as it is not showing growth, and has signs of contamination. We received most of the lab equipment we have ordered </p>
+
<p> Flask 2 discarded as it is not showing growth, and has signs of contamination. We received most of the lab equipment we have ordered. </p>
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+
 
<p> <b> 27/07/15 </b> </p>
 
<p> <b> 27/07/15 </b> </p>
 
BG-11 order arrived finally!
 
BG-11 order arrived finally!
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<p> All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light.</p>
 
<p> All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light.</p>
  
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<h5>What should this page have?</h5>
 
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<ul>
 
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<li>Chronological notes of what your team is doing.</li>
 
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<li> Brief descriptions of  daily important events.</li>
 
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<li>Pictures of your progress. </li>
 
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<li>Mention who participated in what task.</li>
 
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</ul>
 
  
 
</div>
 
</div>
 
</html>
 
</html>

Revision as of 15:12, 28 July 2015

Lab notes

This page is an account of the daily events in our lab and project. Dates are in the UK format DD/MM/YY. 6803 refers to the cyanobacterium used - Synechocystis sp. PCC 6803

29/06/15

  • Cultured a sterile 500ml conical flask using 100 ml BG-11 and 5 loopfuls of 6803 Wild Type (WT). (Flask 1)
  • Cultured 5 streak plates of 6803 WT on BG-11 Agar

    • All placed on the windowsill in our lab under natural light and at room temperature (~25°C)

    02/07/15

  • Three new sterile 500ml conical flasks set up on the shaker at 58rpm and 30°C. Each contains 5 loopfuls of 6803 WT and 100ml of BG-11. They are exposed to the overhead lights in the lab 24/7. (Flasks 2, 3, 4)

    • 07/07/15

  • Flask 5 cultured containing 10ul 6803 WT in 100ml BG-11. This flask will be used for preliminary growth curve measurements. The flask was placed on the shaker with flasks 2,3 and 4 all at 58rpm and 30°C under lab overhead lights.

    • 10/07/15

    2 Litres of water collected from Whiteknights Lake on campus for SODIS, method outlines in protocols.

    13/07/15

    Estimating the Cells per ml for the flask from 29/06 1 a.u at 750nm = 1.6x108

  • OD750 = 0.028
    • Therefore
  • 0.028x1.6x108 = 4.48x106 cells per ml

    • OD750 measurement and cells per ml estimated for flasks cultured on 02/07

    14/07/15

    The lamp safety testing completed so flasks placed under the bright white light bulb

    21/07/15

    Flasks are getting very green, definitely a good sign!

    23/07/15

    Flask 2 discarded as it is not showing growth, and has signs of contamination. We received most of the lab equipment we have ordered.

    27/07/15

    BG-11 order arrived finally!

    1. Miniprep of plasmid DNA: We carried out extraction of biobrick construct plasmids from E.coli NEB-5α glycerol stocks.

      We extracted pSB1K3 plasmid backbones combined with biobricks containing the following genes:

      • PilA1
      • PsaD
      • PetF
      • PilT

      This gave us 4 samples of plasmid DNA, which were placed in the freezer at -20°C.

    2. BG-11 cultures: We added ~200ml of BG-11 to two sterile 500ml conical flasks. We inoculated them as follows;
      • Flask 6 - 1ml of Wild type 6803 from flask 3 as this flask had the best growth.
      • Flask 7 - 10µl of stock Howe Lab triple mutant 6803.
    3. SODIS lakewater cultures: We added ~200ml of SODIS lakewater to two sterile 500ml conical flasks. We inoculated them as follows;
      • SODIS Flask 1 - 1ml of Wild type 6803 from flask 3 as this flask had the best growth.
      • SODIS Flask 2 - 1ml of stock Howe Lab triple mutant 6803.

    All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light.