Difference between revisions of "Team:Reading/Notebook"

Line 26: Line 26:
 
<tr> <td> 20/07 </td> <td> 0.364 </td> <td> 5.824x10<sup>7</sup> </td> </tr>
 
<tr> <td> 20/07 </td> <td> 0.364 </td> <td> 5.824x10<sup>7</sup> </td> </tr>
 
<tr> <td> 21/07 </td> <td> 0.510 </td> <td> 8.160x10<sup>7</sup> </td> </tr>
 
<tr> <td> 21/07 </td> <td> 0.510 </td> <td> 8.160x10<sup>7</sup> </td> </tr>
 +
<tr> <td> 22/07 </td> <td> 0.820 </td> <td> 1.312x10<sup>8</sup> </td> </tr>
 +
<tr> <td> 23/07 </td> <td> 1.016 </td> <td> 1.626x10<sup>8</sup> </td> </tr>
 +
<tr> <td> 24/07 </td> <td> 1.167 </td> <td> 1.876x10<sup>8</sup> </td> </tr>
 +
<tr> <td> 27/07 </td> <td> 1.412 </td> <td> 2.259x10<sup>8</sup> </td> </tr>
 
</table>
 
</table>
 
<p> <b> 10/07/15 </b> </p>  
 
<p> <b> 10/07/15 </b> </p>  
Line 60: Line 64:
 
<tr> <td rowspan="2"> 23/07 </td> <td> 3 </td> <td> 1.302 </td> <td> 2.083x10<sup>8</sup> </td> </tr>
 
<tr> <td rowspan="2"> 23/07 </td> <td> 3 </td> <td> 1.302 </td> <td> 2.083x10<sup>8</sup> </td> </tr>
 
<tr> <td> 4 </td> <td> 1.203 </td> <td> 1.925x10<sup>8</sup> </td> </tr>
 
<tr> <td> 4 </td> <td> 1.203 </td> <td> 1.925x10<sup>8</sup> </td> </tr>
 +
<tr> <td rowspan="2"> 24/07 </td> <td> 3 </td> <td> 1.324 </td> <td> 2.118x10<sup>8</sup> </td> </tr>
 +
<tr> <td> 4 </td> <td> 1.296 </td> <td> 2.074x10<sup>8</sup> </td> </tr>
 +
<tr> <td rowspan="2"> 27/07 </td> <td> 3 </td> <td> 1.691 </td> <td> 2.706x10<sup>8</sup> </td> </tr>
 +
<tr> <td> 4 </td> <td> 1.650 </td> <td> 2.640x10<sup>8</sup> </td> </tr>
 
</table>
 
</table>
 
<p> <b> 14/07/15 </b> </p>
 
<p> <b> 14/07/15 </b> </p>

Revision as of 15:06, 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.
    • Date OD750 Estimated Cells per ml
    07/07 0.000 0
    08/07 0.003 4.8x105
    09/07 0.005 8x105
    10/07 0.005 8x105
    13/07 0.005 8x105
    14/07 0.112 1.792x107
    15/07 0.008 1.28x106
    16/07 0.009 1.44x106
    17/07 0.030 4.8x106
    20/07 0.364 5.824x107
    21/07 0.510 8.160x107
    22/07 0.820 1.312x108
    23/07 1.016 1.626x108
    24/07 1.167 1.876x108
    27/07 1.412 2.259x108

    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

    Date Flask No. OD750 Estimated Cells per ml
    13/07 2 0.006 9.6x105
    3 0.012 1.92x106
    4 0.015 2.4x106
    14/07 2 0.006 9.6x105
    3 0.018 2.88x106
    4 0.017 2.72x106
    15/07 2 0.006 9.6x105
    3 0.021 3.36x106
    4 0.018 2.88x106
    16/07 2 0.001 1.6x105
    3 0.020 3.2x106
    4 0.019 3.04x106
    17/07 2 0.001 1.6x105
    3 0.040 6.4x106
    4 0.027 4.32x106
    20/07 2 0.005 8x105
    3 0.577 9.232x107
    4 0.486 7.776x107
    21/07 2 0.001 1.6x105
    3 0.757 1.211x108
    4 0.653 1.045x108
    22/07 2 0.001 1.6x105
    3 1.049 1.678x108
    4 0.916 1.466x108
    23/07 3 1.302 2.083x108
    4 1.203 1.925x108
    24/07 3 1.324 2.118x108
    4 1.296 2.074x108
    27/07 3 1.691 2.706x108
    4 1.650 2.640x108

    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.

    What should this page have?
    • Chronological notes of what your team is doing.
    • Brief descriptions of daily important events.
    • Pictures of your progress.
    • Mention who participated in what task.