Difference between revisions of "Team:Paris Bettencourt/Notebook/Manufacturing"
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<li>Measure the OD, doing the blank with osmosed water</li> | <li>Measure the OD, doing the blank with osmosed water</li> | ||
<li>If the OD is too high to be measured, make a 10 times dilution and measure the OD of the dilution</li> | <li>If the OD is too high to be measured, make a 10 times dilution and measure the OD of the dilution</li> | ||
− | <li>In the eppendorf tubes, make 10 | + | <li>In the eppendorf tubes, make 10<sup>-1</sup> to 10<sup>-5</sup> dilutions of the yeasts/water solution to plate them on YPDagar+GEN(200)</li></ul> |
<br> | <br> | ||
That is what we did today, and we plated the different dilutions, thanks to the beads. | That is what we did today, and we plated the different dilutions, thanks to the beads. | ||
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<H2>Result of the OD test made on the 27th of July:</h2> | <H2>Result of the OD test made on the 27th of July:</h2> | ||
The 10ml solution of Sc. mcherry/water had an OD of 2,557.<br> | The 10ml solution of Sc. mcherry/water had an OD of 2,557.<br> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-4</sup> dilution grew 359 colonies, so:<br> |
− | <b>1 OD = 1,4x10 | + | <b>1 OD = 1,4x10<sup>7</sup> cells/ml</b><br> |
We repeated the very same OD experiment today, for more repetitions. | We repeated the very same OD experiment today, for more repetitions. | ||
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<h2>Procedure</h2> | <h2>Procedure</h2> | ||
Today, we are repeating the drying experiment (see July 28th) but drying the yeasts on a plastic dish.<br> | Today, we are repeating the drying experiment (see July 28th) but drying the yeasts on a plastic dish.<br> | ||
− | <ul><li>Take 100µL of the culture solution of Sc. mcherry to make dilutions (10 | + | <ul><li>Take 100µL of the culture solution of Sc. mcherry to make dilutions (10<sup>-1</sup> to 10<sup>-5</sup>)</li> |
We remind that the culture tube contain 10ml of media. | We remind that the culture tube contain 10ml of media. | ||
<li>Plate 100µL of each dilutions to deduce the number of cells in the culture tube</li> | <li>Plate 100µL of each dilutions to deduce the number of cells in the culture tube</li> | ||
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<li>Scratch the aluminium to detach the dried yeasts</li> | <li>Scratch the aluminium to detach the dried yeasts</li> | ||
<li>Put the powder in 10ml of osmosed water</li> | <li>Put the powder in 10ml of osmosed water</li> | ||
− | <li>Take 100µL of the solution to make dilutions (10 | + | <li>Take 100µL of the solution to make dilutions (10<sup>-1</sup> to 10<sup>-5</sup>)</li> |
<li>Thanks to the beads, plate 100µL of each dilution</li></ul> | <li>Thanks to the beads, plate 100µL of each dilution</li></ul> | ||
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<h2>Result of the OD test made on the 29th of July:</h2> | <h2>Result of the OD test made on the 29th of July:</h2> | ||
The 10ml solution of Sc. mcherry/water had an OD of 2,635.<br> | The 10ml solution of Sc. mcherry/water had an OD of 2,635.<br> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-4</sup> dilution grew 205 colonies, so:<br> |
− | <b>1 OD = 7,8x10 | + | <b>1 OD = 7,8x10<sup>6</sup> cells/ml</b> |
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<h1 class="date two">August 1st</h1> | <h1 class="date two">August 1st</h1> | ||
<h2>Result of the drying test made on the 30th of July:</h2> | <h2>Result of the drying test made on the 30th of July:</h2> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-5</sup> dilution of the initial 10ml culture solution grew 106 colonies, so we initially dried 1,06x10<sup>9</sup> cells.<br> |
− | 100µL of the 10 | + | 100µL of the 10<sup>-4</sup> dilution of the powder mixed with 30ml of osmosed water grew 123 colonies, so 8,4x10<sup>8</sup> cells survived.<br> |
We therefore have a survival rate of 79%. | We therefore have a survival rate of 79%. | ||
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<br> | <br> | ||
Our second idea was to cook a homemade powder with edible ingredients easily found in India.<br> | Our second idea was to cook a homemade powder with edible ingredients easily found in India.<br> | ||
− | We found a recipe that inspired us | + | We found a recipe that inspired us <a href="www.instructables.com">here</a>. <br> |
We decided to compare the initial recipe to our modifications, so we made for different tests :<br> | We decided to compare the initial recipe to our modifications, so we made for different tests :<br> | ||
<ul> | <ul> | ||
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We tought it could be a good media of distribution.<br><br> | We tought it could be a good media of distribution.<br><br> | ||
We dissolved one cube in 10ml of osmosed water, after weighed it.<br> | We dissolved one cube in 10ml of osmosed water, after weighed it.<br> | ||
− | Then we made dilutions (10 | + | Then we made dilutions (10<sup>-1</sup> to 10<sup>-5</sup>) and plated 100µL of each on YPDagar+GEN(200) |
</p></div> | </p></div> | ||
<div class="column-right"><p> | <div class="column-right"><p> | ||
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<li>Measure the OD, doing the blank with osmosed water</li> | <li>Measure the OD, doing the blank with osmosed water</li> | ||
<li>If the OD is too high to be measured, make a 10 times dilution and measure the OD of the dilution</li> | <li>If the OD is too high to be measured, make a 10 times dilution and measure the OD of the dilution</li> | ||
− | <li>In the eppendorf tubes, make 10 | + | <li>In the eppendorf tubes, make 10<sup>-1</sup> to 10<sup>-5</sup> dilutions of the G1513/water solution to plate 100µL of each on M17Glucose+agar+Ery(10)</li></ul> |
<br> | <br> | ||
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</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-1</sup></td> |
<td>381</td> | <td>381</td> | ||
</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-2</sup></td> |
<td>84</td> | <td>84</td> | ||
</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10<sup-3</sup</td> | + | <td>10<sup>-3</sup></td> |
<td>11</td> | <td>11</td> | ||
</tr> | </tr> | ||
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We have, in average, 7,74x10<sup>3</sup> cells per ml of the 10<sup>-1</sup> dilution.<br> | We have, in average, 7,74x10<sup>3</sup> cells per ml of the 10<sup>-1</sup> dilution.<br> | ||
− | so: <b>1 OD = 6,0x10 | + | so: <b>1 OD = 6,0x10<sup>4</sup> cells/ml</b><br> |
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
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For Sc. mcherry:<br> | For Sc. mcherry:<br> | ||
The 10ml solution of Sc. mcherry/water had an OD of 1,820.<br> | The 10ml solution of Sc. mcherry/water had an OD of 1,820.<br> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-4</sup> dilution grew 70 colonies, so:<br> |
− | <b>1 OD = 3,8x10 | + | <b>1 OD = 3,8x10<sup>6</sup> cells/ml</b><br> |
For G1513:<br> | For G1513:<br> | ||
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</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-5</sup></td> |
<td>131</td> | <td>131</td> | ||
</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-6</sup></td> |
<td>21</td> | <td>21</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | So <b>1 OD = 1,1x10 | + | So <b>1 OD = 1,1x10<sup>8</sup> cells/ml</b> |
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For Sc. mcherry:<br> | For Sc. mcherry:<br> | ||
The 10ml solution of Sc. mcherry/water had an OD of 1,828.<br> | The 10ml solution of Sc. mcherry/water had an OD of 1,828.<br> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-5</sup> dilution grew 56 colonies, so:<br> |
− | <b>1 OD = 3,1x10 | + | <b>1 OD = 3,1x10<sup>6</sup> cells/ml</b><br> |
For G1513:<br> | For G1513:<br> | ||
Nothing... It seems that the colony used was dead so the cubes will not contained any G1513.<br> | Nothing... It seems that the colony used was dead so the cubes will not contained any G1513.<br> | ||
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</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-4</sup></td> |
<td>505</td> | <td>505</td> | ||
</tr> | </tr> | ||
<tr align="center"> | <tr align="center"> | ||
− | <td>10 | + | <td>10<sup>-5</sup></td> |
<td>39</td> | <td>39</td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | <b>1 OD = 2,6x10 | + | <b>1 OD = 2,6x10<sup>6</sup> cells/mL</b><br> |
The second 10ml solution of Sc. mcherry/water had an OD of 2,523.<br> | The second 10ml solution of Sc. mcherry/water had an OD of 2,523.<br> | ||
− | 100µL of the 10 | + | 100µL of the 10<sup>-5</sup> dilution grew 95 colonies.<br> |
so <b>1 OD = ?????????????????? cells/mL</b><br> | so <b>1 OD = ?????????????????? cells/mL</b><br> | ||
For G1513:<br> | For G1513:<br> |
Revision as of 17:19, 14 September 2015