Difference between revisions of "Team:Pasteur Paris/Week 10"
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<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 /> | ||
<u>3 solutions: </u><br /> | <u>3 solutions: </u><br /> | ||
− | + | <br /> | |
<u>Solution 1: (salts)</u><br /> | <u>Solution 1: (salts)</u><br /> | ||
5,3g of Potassium Phosphate (dibasic) K2HPO4<br /> | 5,3g of Potassium Phosphate (dibasic) K2HPO4<br /> | ||
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Autoclave<br /> | Autoclave<br /> | ||
Complete with water to 333 mL</p> | Complete with water to 333 mL</p> | ||
− | <p><u>Solution 2: (agar)</u><br /> | + | <p style="text-align:left"><u>Solution 2: (agar)</u><br /> |
16g of agar<br /> | 16g of agar<br /> | ||
Complete with water to 333 mL<br /> | Complete with water to 333 mL<br /> | ||
Autoclave</p> | Autoclave</p> | ||
− | <p><u>Solution 3: (sugar)</u><br /> | + | <p style="text-align:left"><u>Solution 3: (sugar)</u><br /> |
In fact we have to put 4g of sugar in 333 mL of H2O. But we wanted to see if bacteria took TPA for a carbon source. So just the water was put. Next the flacon was autoclaved.</p> | In fact we have to put 4g of sugar in 333 mL of H2O. But we wanted to see if bacteria took TPA for a carbon source. So just the water was put. Next the flacon was autoclaved.</p> | ||
<p> </p> | <p> </p> | ||
− | <p>When the three parts have been autoclaved, 2 ingredients must be add to the medium in aseptically conditions:<br /> | + | <p style="text-align:left">When the three parts have been autoclaved, 2 ingredients must be add to the medium in aseptically conditions:<br /> |
1mL of 10% Magnesium Sulfate MgSO4 (autoclaved too) (1,5g in 15mL of H2O).<br /> | 1mL of 10% Magnesium Sulfate MgSO4 (autoclaved too) (1,5g in 15mL of H2O).<br /> | ||
1mL of 0,2% Thiamin (vitamin B1).</p> | 1mL of 0,2% Thiamin (vitamin B1).</p> |
Revision as of 00:48, 13 September 2015
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Week 10 Toxicity and solubility of TPA
Preparation of M9 and minimal media Solution 2: (agar) Solution 3: (sugar)
When the three parts have been autoclaved, 2 ingredients must be add to the medium in aseptically conditions: At the end the volume of Agar was too important (350mL and not 333mL). So we had to redo this experience the next day. 05-08-15 – Experiment: Preparation of M9 medium – VB, PV Aim: Get medium for Pfeifer cells culture. Protocol: (for 1L medium) Agar was directly put in a bottle and salts were dissolved in a beaker with water. 6g of Sodium Phosphate Na2HPO4.H2O When this flacon was autoclaved some ingredients were added in aseptically conditions: 1mL of 1M Magnesium sulphate MgSO4 (6,02g in 50mL 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 – VB, PV Aim: Have prepared M9 Medium and Minimal Medium for Petri dishes We restart the two last protocols but this time we included the agar for the future Petri dishes. Solution 1: (salts) Solution 2: (agar) Solution 3: We underestimate the time for agar to solidify, so we had solid agar in an eprouvette. Moreover one of the flacon 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 mediums were ready but they did not have the last ingredients.
Enzymatic Assay of NB-EsteraseFigure 1: Gel de migration pour verification de la ligation. There is no band for the ligation sample. Gibson Assembly
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