Difference between revisions of "Team:Pasteur Paris/Week 10"
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<h3><em>Toxicity and solubility of TPA (Terephthalic acid)</em></h3><br> | <h3><em>Toxicity and solubility of TPA (Terephthalic acid)</em></h3><br> | ||
<ol> | <ol> | ||
− | <li style="text-align:left">Dissolution of TPA in DMSO (best concentration | + | <li style="text-align:left">Dissolution of TPA in DMSO (Dimethyl sulfoxide). The best concentration obtained is 62 mg/l </li> |
<li style="text-align:left">Dilution of TPA stock in LB Broth => TPA precipitates at a concentration of 3.9 g/l, maximum concentration in LB Broth is 0.9 mg/l </li> | <li style="text-align:left">Dilution of TPA stock in LB Broth => TPA precipitates at a concentration of 3.9 g/l, maximum concentration in LB Broth is 0.9 mg/l </li> | ||
<p style="text-align:left"><em>Preparation of M9 and minimal media<u></u></em><br /> | <p style="text-align:left"><em>Preparation of M9 and minimal media<u></u></em><br /> | ||
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1 ml of 0.1 M of Calcium Chloride CaCl<sub>2</sub> (0.55 g in 50 ml of H<sub>2</sub>O)</p> | 1 ml of 0.1 M of Calcium Chloride CaCl<sub>2</sub> (0.55 g in 50 ml of H<sub>2</sub>O)</p> | ||
<p style="text-align:left">A mistake was done yesterday but this time an other did too: The last ingredients were added but they were not sterile so the medium failed again.</p> | <p style="text-align:left">A mistake was done yesterday but this time an other did too: The last ingredients were added but they were not sterile so the medium failed again.</p> | ||
− | <p align="center"><u>06-08-2015 </u><u>–</u><u> </u><u>Experiment: Minimal and M9 medium for TPA Toxicity and BAP</u><u> </u><u>–</u><u> </u><u>VB, PV</u><u> </u><br /> | + | <p align="center"><u>06-08-2015 </u><u>–</u><u> </u><u>Experiment: Minimal and M9 medium for TPA Toxicity and BAP 1</u><u> </u><u>–</u><u> </u><u>VB, PV</u><u> </u><br /> |
<u>Redaction by VB</u><u> </u></p> | <u>Redaction by VB</u><u> </u></p> | ||
<p style="text-align:left"><strong><u>Aim:</u></strong> Have prepared M9 medium and minimal medium for Petri dishes</p> | <p style="text-align:left"><strong><u>Aim:</u></strong> Have prepared M9 medium and minimal medium for Petri dishes</p> | ||
<p style="text-align:left">We restarted the last two protocols but this time we included the agar for the future Petri dishes.<br /> | <p style="text-align:left">We restarted the last two protocols but this time we included the agar for the future Petri dishes.<br /> | ||
− | For the future experiments every medium will have the same concentration of sugar for the bacteria growth, and the test of the TPA toxicity. | + | For the future experiments every medium will have the same concentration of sugar for the bacteria growth, and the test of the TPA toxicity. We began by a theoretical concentration for our Petri dishes: to see the saturation concentration of TPA we decided to do dilutions at 1/10 and 1/20. <br /> |
This time we decided to make 1.5 l of minimal medium<br /> | This time we decided to make 1.5 l of minimal medium<br /> | ||
<strong><u>Protocol:</u></strong><strong><u> </u></strong></p> | <strong><u>Protocol:</u></strong><strong><u> </u></strong></p> | ||
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<tr> | <tr> | ||
<td width="74" valign="center"><p>Biobrick</p></td> | <td width="74" valign="center"><p>Biobrick</p></td> | ||
− | <td width="106" valign="top"><p>DNA Concentration (ng/ | + | <td width="106" valign="top"><p>DNA Concentration (ng/µl)</p></td> |
− | <td width="88" valign="top"><p>OD ( | + | <td width="88" valign="top"><p>OD (230 nm)</p></td> |
− | <td width="88" valign="top"><p>OD ( | + | <td width="88" valign="top"><p>OD (260 nm)</p></td> |
− | <td width="71" valign="top"><p>OD ( | + | <td width="71" valign="top"><p>OD (280 nm)</p></td> |
− | <td width="106" valign="top"><p>OD ( | + | <td width="106" valign="top"><p>OD (260 nm) / OD (280 nm)</p></td> |
− | <td width="109" valign="top"><p>OD ( | + | <td width="109" valign="top"><p>OD (260 nm) / OD (230 nm)</p></td> |
</tr> | </tr> | ||
<tr> | <tr> |
Revision as of 23:52, 17 September 2015
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Week 10 Toxicity and solubility of TPA (Terephthalic acid)
Preparation of M9 and minimal media Solution 2: (agar) Solution 3: (sugar)
When the three parts have been autoclaved, 2 more ingredients must be added to the medium aseptically: At the end the volume of Agar was exceeded (350 ml and not 333 ml). So we had to redo this experiment the next day. 05-08-15 – Experiment: Preparation of M9 medium – VB, PV Aim: Get culture medium for BAP 1 Pfeifer cells. Protocol: (for 1 l medium) Agar was directly put in a bottle and salts were dissolved in a beaker with water. 6 g of Sodium Phosphate Na2HPO4.H2O When this flask was autoclaved some ingredients were added in aseptical conditions: 1 ml of 1M Magnesium sulphate MgSO4 (6.02 g in 50 ml H2O) A mistake was done yesterday but this time an other did too: The last ingredients were added but they were not sterile so the medium failed again. 06-08-2015 – Experiment: Minimal and M9 medium for TPA Toxicity and BAP 1 – VB, PV Aim: Have prepared M9 medium and minimal medium for Petri dishes We restarted the last two protocols but this time we included the agar for the future Petri dishes. Solution 1: (salts) Solution 2: (agar) Solution 3: We underestimated the time for agar to solidify, so we had solid agar in the test-tube. Moreover one of the flask was not sterile so we contaminated our medium. We decided to return at the first protocol (for 1L). At the end of this journey the two media were ready but they did not have the last ingredients.
Enzymatic Assay of NB-EsteraseFigure 8: Gel for verification of ligation. There is no band for the ligation sample. Gibson Assembly
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