Difference between revisions of "Team:Goettingen/Notebook"

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<div class="notebook-item">
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    <div class="notebook-head">
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        <h3 class="notebook-head_title">Used recipes for buffers and media
 +
        </h3>
 +
        <p class="notebook-head_date">25.04.18</p>
 +
    </div>
 +
    <div class="notebook-content">
 +
        <div class="notebook-table article_table">
 +
            <p>LB-Medium</p>
 +
            <table>
 +
                <tr>
 +
                    <th>Components</th>
 +
                    <th class="article-table_right">Amount (g)</th>
 +
                </tr>
 +
                <tr>
 +
                    <td>Tryptone</td>
 +
                    <td class="article-table_right">10</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>NaCl</td>
 +
                    <td class="article-table_right">10</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>Yeast exctract</td>
 +
                    <td class="article-table_right">5</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>1000 mL H<sub>2</sub>O</td>
 +
                    <td></td>
 +
                </tr>
 +
                <tr class="article_table_hline">
 +
                    <td><strong>Autoclavation</strong></td>
 +
                    <td></td>
 +
                </tr>
 +
            </table>
 +
        </div>
 +
        <div class="notebook-table article_table">
 +
            <p>SP-Medium</p>
 +
            <table>
 +
                <tr>
 +
                    <th>Components</th>
 +
                    <th class="article-table_right">Amount (g)</th>
 +
                </tr>
 +
                <tr>
 +
                    <td>Nutrient broth</td>
 +
                    <td class="article-table_right">8</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>MgS0<sub>4</sub> &#9679; 7 H<sub>2</sub>O</td>
 +
                    <td class="article-table_right">10</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>KCl</td>
 +
                    <td class="article-table_right">1</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>1000 mL H<sub>2</sub>O</td>
 +
                    <td></td>
 +
                </tr>
 +
                <tr>
 +
                    <td><strong>Autoclavation</strong></td>
 +
                    <td></td>
 +
                </tr>
 +
                <tr>
 +
                    <td>CaCl<sub>2</sub></td>
 +
                    <td class="article-table_right">0.5&nbsp;M</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>MnCl<sub>2</sub></td>
 +
                    <td class="article-table_right">10&nbsp;mM</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>Iron ferric ammonium citrate</td>
 +
                    <td class="article-table_right">2&nbsp;mL</td>
 +
                </tr>
 +
 
 +
            </table>
 +
        </div>
 +
        <div class="notebook-table article_table">
 +
            <p>10&times; MN-Medium</p>
 +
            <table>
 +
                <tr>
 +
                    <th>Components</th>
 +
                    <th class="article-table_right">Amount (g)</th>
 +
                </tr>
 +
                <tr>
 +
                    <td>K<sub>2</sub>HPO<sub>4</sub> &#9679; 3 H<sub>2</sub>O</td>
 +
                    <td class="article-table_right">136</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>KH<sub>2</sub>PO<sub>4</sub></td>
 +
                    <td class="article-table_right">60</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>Na citrate</td>
 +
                    <td class="article-table_right">10</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>1000 mL H<sub>2</sub>O</td>
 +
                    <td></td>
 +
                </tr>
 +
                <tr class="article_table_hline">
 +
                    <td><strong>Ad to 50 mL with sterile water</strong></td>
 +
                    <td class="article-table_right">100</td>
 +
                </tr>
 +
            </table>
 +
        </div>
 +
        <div class="notebook-table article_table">
 +
            <p>CS-Glucose Medium</p>
 +
            <table>
 +
            <tr>
 +
                <th>Components</th>
 +
                <th class="article-table_right">Amount (ml)</th>
 +
            </tr>
 +
                <tr>
 +
                    <td>C-Salts</td>
 +
                    <td class="article-table_right">10</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>III-Salts</td>
 +
                    <td class="article-table_right">0.5</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>Tryptophane</td>
 +
                    <td class="article-table_right">0.5</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>Na-Succinate</td>
 +
                    <td class="article-table_right">1</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>50 % Glucose</td>
 +
                    <td class="article-table_right">0.5</td>
 +
                </tr>
 +
                <tr class="article_table_hline">
 +
                    <td><strong>Ad to 50 mL with sterile water</strong></td>
 +
            <td></td>
 +
            </table>
 +
        </div>
 +
       
 +
            <div class="notebook-table article_table">
 +
            <p>CS-Glucose Agar Plates</p>
 +
            <table>
 +
            <tr>
 +
                <th>Components</th>
 +
                <th class="article-table_right">Amount (ml)</th>
 +
            </tr>
 +
                <tr>
 +
                    <td>CS-Glucose Medium</td>
 +
                    <td class="article-table_right">25</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>3 % Bacto-Agar</td>
 +
                    <td class="article-table_right">25</td>
 +
                </tr>
 +
                </table>
 +
        </div>
 +
    </div>
 +
  </div>
 +
<div class="notebook-item">
 +
    <div class="notebook-head">
 +
        <h3 class="notebook-head_title"></h3>
 +
        <p class="notebook-head_date">27.04.18</p>
 +
    </div>
 +
    <div class="notebook-content">
 +
        <p>Cryo cultures of strains 168, BP233, BP234 were prepared with 90% LB medium and 10% DMSO. The cells were frozen and stored at -80°C.</p>
 +
    </div>
 +
</div>
 +
<div class="notebook-item">
 +
    <div class="notebook-head">
 +
        <h3 class="notebook-head_title"></h3>
 +
        <p class="notebook-head_date">02.05.18</p>
 +
    </div>
 +
    <div class="notebook-content">
 +
        <p>The strains BP233, BP235 and the WT strain 168 were propagated to 3 ml LB medium and inoculated for 4 h at 37°C and 220 rpm. They were harvested in a 2 ml reaction tube by centrifugation at 13000 rpm for one minute. The supernatant was discared and the cells were washed in 1x C-Salts. Afterwards, they were resuspended in 300 µl 1x C-Salts. The OD<sub>600</sub> was determined.</p>
 +
                    <div class="notebook-table article_table">
 +
            <table>
 +
                <tr>
 +
                    <th>Strain</th>
 +
                    <th class="article-table_right">OD<sub>600</sub></th>
 +
                </tr>
 +
                <tr>
 +
                    <td>168</td>
 +
                    <td class="article-table_right">9.8</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>BP233</td>
 +
                    <td class="article-table_right">6.1</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>BP235</td>
 +
                    <td class="article-table_right">7.6</td>
 +
                </tr>
 +
            </table>
 +
    </div>
 +
 
 +
        <p>The cells were then struck out on cs-glucose agar plates with variating glyphosate concentrations. 168 was propagated on plates containing 0 mM  and 10 mM glyphosate, while BP233 was additionally propagated on plates with the glyphosate concentrations of 30 mM and 40 mM.BP235 was additonally propagated on plates containing glyphosate concentrations of 50 mM and 60 mM. All agar plates were then inoculated at 37 °C for 2 days.  </p>
 +
        <p>The strains 168, BP233 and BP236 were propagated to 3 ml LB medium and inoculated for 4 h at 37°C and 220 rpm. They were harvested in a 2 ml reaction tube by centrifugation at 13000 rpm for one minute. The supernatant was discared and the cells were washed in 1x C-Salts. Afterwards, they were resuspended in 300 µl 1x C-Salts. The OD<sub>600</sub> was determined.</p>
 +
              <div class="notebook-table article_table">
 +
            <table>
 +
                <tr>
 +
                    <th>Strain</th>
 +
                    <th class="article-table_right">OD<sub>600</sub></th>
 +
                </tr>
 +
                <tr>
 +
                    <td>168</td>
 +
                    <td class="article-table_right">10.74</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>BP233</td>
 +
                    <td class="article-table_right">8.37</td>
 +
                </tr>
 +
                <tr>
 +
                    <td>BP236</td>
 +
                    <td class="article-table_right">7.71</td>
 +
                </tr>
 +
            </table>
 +
    </div>
 +
 
 +
        <p>100 µl of solutions were propagated on CS-Glucose agar plates containing 0 mM glyphosate as well as plates containing 10 mM glyphosate. These plates were inoculated at 37 °C for 2 days.</p>
 +
 
 +
</div>
 +
</div>
 +
<div class="notebook-item">
 +
    <div class="notebook-head">
 +
        <h3 class="notebook-head_title"></h3>
 +
        <p class="notebook-head_date">04.05.18</p>
 +
    </div>
 +
    <div class="notebook-content">
 +
        <p>The growth of the evolved suppressor mutants of the strains 168, BP233 and BP235 on 10 mM and 0 mM glyphosate CS-glucose agar plates was documented.</p>
 +
        <p>A growth test of 168, BP233 and BP237 on CS-Glucose agar plates containing 0 mM and 10 mM glyphosate.</p>
 +
    </div>
 +
</div>
 +
</body>
 
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Revision as of 14:41, 14 September 2018

Our Labnotes:

April

25.04.18 – 30.04.18

Used recipes for buffers and media

25.04.18

LB-Medium

Components Amount (g)
Tryptone 10
NaCl 10
Yeast exctract 5
1000 mL H2O
Autoclavation

SP-Medium

Components Amount (g)
Nutrient broth 8
MgS04 ● 7 H2O 10
KCl 1
1000 mL H2O
Autoclavation
CaCl2 0.5 M
MnCl2 10 mM
Iron ferric ammonium citrate 2 mL

10× MN-Medium

Components Amount (g)
K2HPO4 ● 3 H2O 136
KH2PO4 60
Na citrate 10
1000 mL H2O
Ad to 50 mL with sterile water 100

CS-Glucose Medium

Components Amount (ml)
C-Salts 10
III-Salts 0.5
Tryptophane 0.5
Na-Succinate 1
50 % Glucose 0.5
Ad to 50 mL with sterile water

CS-Glucose Agar Plates

Components Amount (ml)
CS-Glucose Medium 25
3 % Bacto-Agar 25

27.04.18

Cryo cultures of strains 168, BP233, BP234 were prepared with 90% LB medium and 10% DMSO. The cells were frozen and stored at -80°C.

02.05.18

The strains BP233, BP235 and the WT strain 168 were propagated to 3 ml LB medium and inoculated for 4 h at 37°C and 220 rpm. They were harvested in a 2 ml reaction tube by centrifugation at 13000 rpm for one minute. The supernatant was discared and the cells were washed in 1x C-Salts. Afterwards, they were resuspended in 300 µl 1x C-Salts. The OD600 was determined.

Strain OD600
168 9.8
BP233 6.1
BP235 7.6

The cells were then struck out on cs-glucose agar plates with variating glyphosate concentrations. 168 was propagated on plates containing 0 mM and 10 mM glyphosate, while BP233 was additionally propagated on plates with the glyphosate concentrations of 30 mM and 40 mM.BP235 was additonally propagated on plates containing glyphosate concentrations of 50 mM and 60 mM. All agar plates were then inoculated at 37 °C for 2 days.

The strains 168, BP233 and BP236 were propagated to 3 ml LB medium and inoculated for 4 h at 37°C and 220 rpm. They were harvested in a 2 ml reaction tube by centrifugation at 13000 rpm for one minute. The supernatant was discared and the cells were washed in 1x C-Salts. Afterwards, they were resuspended in 300 µl 1x C-Salts. The OD600 was determined.

Strain OD600
168 10.74
BP233 8.37
BP236 7.71

100 µl of solutions were propagated on CS-Glucose agar plates containing 0 mM glyphosate as well as plates containing 10 mM glyphosate. These plates were inoculated at 37 °C for 2 days.

04.05.18

The growth of the evolved suppressor mutants of the strains 168, BP233 and BP235 on 10 mM and 0 mM glyphosate CS-glucose agar plates was documented.

A growth test of 168, BP233 and BP237 on CS-Glucose agar plates containing 0 mM and 10 mM glyphosate.