Difference between revisions of "Team:Paris Bettencourt/Notebook/Manufacturing"
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{{Paris_Bettencourt/header}} | {{Paris_Bettencourt/header}} | ||
{{Paris_Bettencourt/menu}} | {{Paris_Bettencourt/menu}} | ||
− | {{Paris_Bettencourt/ | + | {{Paris_Bettencourt/banner|page_id=notebook|page_name=Notebook - Manufacturing}} |
<html> | <html> | ||
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The goal of this project is to create an way to grow and distribute our strains in India, easily and for cheap.<br> | The goal of this project is to create an way to grow and distribute our strains in India, easily and for cheap.<br> | ||
Since our genetically engineered strains will be ready only in september, we will not have time to work on it, so we have to find similar strains, that we can study and select with antibiotics.<br> | Since our genetically engineered strains will be ready only in september, we will not have time to work on it, so we have to find similar strains, that we can study and select with antibiotics.<br> | ||
− | We chose Saccharomyces cerevisiae mcherry, wich is resistant to geneticin at a concentration of 200µg/mL, and produce RFP.<br> | + | We chose Saccharomyces cerevisiae mcherry, wich is resistant to geneticin at a concentration of 200µg/mL, and produce RFP. Therefore, it's easy to select and characterize it.<br> |
Basically, we are going to try different ways to distribute the strains and see how well they survived for each process.<br> | Basically, we are going to try different ways to distribute the strains and see how well they survived for each process.<br> | ||
In order to do that, we will have to calculate a survival rate so we have to be able to know how many cells we are putting in the beggining, in the recipe we are trying. | In order to do that, we will have to calculate a survival rate so we have to be able to know how many cells we are putting in the beggining, in the recipe we are trying. | ||
Line 97: | Line 97: | ||
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<sup>-4</sup> dilution grew 359 colonies, so:<br> | 100µL of the 10<sup>-4</sup> dilution grew 359 colonies, so:<br> | ||
− | <b>1 | + | <b>1 A = 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. | ||
Line 129: | Line 129: | ||
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<sup>-4</sup> dilution grew 205 colonies, so:<br> | 100µL of the 10<sup>-4</sup> dilution grew 205 colonies, so:<br> | ||
− | <b>1 | + | <b>1 A = 7,8x10<sup>6</sup> cells/ml</b> |
− | + | <br> | |
<a name="august" class="anchor"><h1></h1></a> | <a name="august" class="anchor"><h1></h1></a> | ||
<h1 class="date two">August 1st</h1> | <h1 class="date two">August 1st</h1> | ||
Line 145: | Line 145: | ||
<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 <a href="www.instructables.com">here</a>. <br> | + | We found a recipe that inspired us <a href="http://www.instructables.com/id/Stop-Paying-for-Yeast-Make-Your-Own/">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> | ||
Line 242: | Line 242: | ||
Survival rate: | Survival rate: | ||
<ul> | <ul> | ||
− | <li>Wheat flour, sugar and ginger: | + | <li>Wheat flour, sugar and ginger: 14% </li> |
− | <li>Rice flour, sugar and ginger: | + | <li>Rice flour, sugar and ginger: 20%</li> |
− | <li>Rice flour, sugar: | + | <li>Rice flour, sugar: 12%</li> |
− | <li>Rice flour: | + | <li>Rice flour: 4%</li> |
</ul> | </ul> | ||
<h2>Discussion:</h2> | <h2>Discussion:</h2> | ||
Line 257: | Line 257: | ||
<div class="column-left"><p> | <div class="column-left"><p> | ||
<br> | <br> | ||
− | We obtained a survival rate of | + | We obtained a survival rate of 69%. <br><br> |
After staying 5hours in the water, the survival rate reached 737%. | After staying 5hours in the water, the survival rate reached 737%. | ||
</p></div> | </p></div> | ||
Line 319: | Line 319: | ||
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 | + | so: <b>1 A = 6,0x10<sup>4</sup> cells/ml</b><br> |
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
Line 399: | Line 399: | ||
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<sup>-4</sup> dilution grew 70 colonies, so:<br> | 100µL of the 10<sup>-4</sup> dilution grew 70 colonies, so:<br> | ||
− | <b>1 | + | <b>1 A = 3,8x10<sup>6</sup> cells/ml</b><br> |
For G1513:<br> | For G1513:<br> | ||
Line 418: | Line 418: | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | So <b>1 | + | So <b>1 A = 1,1x10<sup>8</sup> cells/ml</b> |
Line 449: | Line 449: | ||
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<sup>-5</sup> dilution grew 56 colonies, so:<br> | 100µL of the 10<sup>-5</sup> dilution grew 56 colonies, so:<br> | ||
− | <b>1 | + | <b>1 A = 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> | ||
Line 479: | Line 479: | ||
</tr> | </tr> | ||
</table> | </table> | ||
− | <b>1 | + | <b>1 A = 2,3x10<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<sup>-5</sup> dilution grew 95 colonies.<br> | 100µL of the 10<sup>-5</sup> dilution grew 95 colonies.<br> | ||
− | so <b>1 | + | so <b>1 A = 3,8x10<sup>6</sup> cells/mL</b><br> |
For G1513:<br> | For G1513:<br> | ||
Nothing... We used cells from the previous culture, wich was dead, so it seems normal. | Nothing... We used cells from the previous culture, wich was dead, so it seems normal. | ||
Line 524: | Line 524: | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
We want to see how the strains in the cubes react when put in idli.<br> | We want to see how the strains in the cubes react when put in idli.<br> | ||
− | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
<ul> | <ul> | ||
Line 531: | Line 530: | ||
<li>Dilute the cube in 5ml of osmosed water</li> | <li>Dilute the cube in 5ml of osmosed water</li> | ||
<li>Add this solution to 1 bottle of idli before the fermentation</li> | <li>Add this solution to 1 bottle of idli before the fermentation</li> | ||
+ | <li>After the fermentation, plate 100μL of a 10<sup>-2</sup> dilution of the Idli water</li> | ||
</ul> | </ul> | ||
Line 541: | Line 541: | ||
<h2>Results of the idli test</h2> | <h2>Results of the idli test</h2> | ||
− | + | <table style="width:100%; text-align:center"> | |
+ | <tr> | ||
+ | <th></th> | ||
+ | <th>Number of cells in the top part of the idli, 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of cells in the middle part of the idli, 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of cells in the bottom part of the idli, 10<sup>-2</sup> dilution</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>X</th> | ||
+ | <td>38 / 23</td> | ||
+ | <td>324</td> | ||
+ | <td>71 / 111</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>Y</th> | ||
+ | <td>0</td> | ||
+ | <td>39 / 27</td> | ||
+ | <td>64 / 47</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <br> | ||
− | <a name="september" class="anchor"> | + | <a name="september" class="anchor"><h1 class="date three">September 2nd</h1></a> |
− | + | ||
− | <h1 class="date three">September 2nd</h1> | + | |
<h2>Results of the second media test made on the 31st of August</h2> | <h2>Results of the second media test made on the 31st of August</h2> | ||
It's a fail, there was almost only water in the well so nothing grew.<br> | It's a fail, there was almost only water in the well so nothing grew.<br> | ||
Line 574: | Line 592: | ||
After centrifugation, throwing the media and replacing it by osmosed water, the cultures gave us two solutions: | After centrifugation, throwing the media and replacing it by osmosed water, the cultures gave us two solutions: | ||
<ul><li>Solution X: 8,8x10<sup>7</sup> cells of Sc. mcherry per ml</li> | <ul><li>Solution X: 8,8x10<sup>7</sup> cells of Sc. mcherry per ml</li> | ||
− | <li>Solution Y: 5,2x10<sup>9</sup> cells of G1513 per ml</li> | + | <li>Solution Y: 5,2x10<sup>9</sup> cells of G1513 per ml</li></ul> |
We made 3 different types of cubes: | We made 3 different types of cubes: | ||
Line 645: | Line 663: | ||
<h1 class="date three">September 9th</h1> | <h1 class="date three">September 9th</h1> | ||
We plated the first cubes today.<br> | We plated the first cubes today.<br> | ||
− | Everyday, we will follow the same protocol | + | Everyday, we will follow the same protocol: |
+ | <ul> | ||
+ | <li>Weigh the cube</li> | ||
+ | <li>Dilute the cube in 10ml of osmosed water during 5min</li> | ||
+ | <li>Dilute the solution obtained 10<sup>-1</sup>, 10<sup>-2</sup> and 10<sup>-3</sup> times</li> | ||
+ | <li>Plate the dilutions</li> | ||
+ | </ul> | ||
+ | This should only take 20min in total, doing it for all the cubes in the same time.<br> | ||
For more clarity, we will write the results for each cubes on the day we plated it (even if the colonies took 1 or 2 days to grow depending on the strains, so we only add the results 2 days after the plating). | For more clarity, we will write the results for each cubes on the day we plated it (even if the colonies took 1 or 2 days to grow depending on the strains, so we only add the results 2 days after the plating). | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 663: | Line 688: | ||
<td>72</td> | <td>72</td> | ||
<td>10</td> | <td>10</td> | ||
− | <td></td> | + | <td>8,6x10<sup>6</sup></td> |
− | <td></td> | + | <td>136,5%</td> |
</tr> | </tr> | ||
Line 673: | Line 698: | ||
<td>80</td> | <td>80</td> | ||
<td>11</td> | <td>11</td> | ||
− | <td></td> | + | <td>9,5x10<sup>6</sup></td> |
− | <td></td> | + | <td>166,7%</td> |
</tr> | </tr> | ||
Line 683: | Line 708: | ||
<td>79</td> | <td>79</td> | ||
<td>10</td> | <td>10</td> | ||
− | <td></td> | + | <td>9,0x10<sup>6</sup></td> |
− | <td></td> | + | <td>2,4%</td> |
</tr> | </tr> | ||
Line 693: | Line 718: | ||
<td>357</td> | <td>357</td> | ||
<td>23</td> | <td>23</td> | ||
− | <td></td> | + | <td>2,9x10<sup>7</sup></td> |
− | <td></td> | + | <td>5,4%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 700: | Line 725: | ||
<br> | <br> | ||
<h1 class="date three">September 10th</h1> | <h1 class="date three">September 10th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 714: | Line 739: | ||
<td>0,97</td> | <td>0,97</td> | ||
<td>6,4x10<sup>6</sup></td> | <td>6,4x10<sup>6</sup></td> | ||
− | <td> | + | <td>104</td> |
− | <td> | + | <td>5</td> |
− | <td></td> | + | <td>7,7x10<sup>6</sup></td> |
− | <td></td> | + | <td>120,3</td> |
</tr> | </tr> | ||
Line 724: | Line 749: | ||
<td>1,40</td> | <td>1,40</td> | ||
<td>8,3x10<sup>6</sup></td> | <td>8,3x10<sup>6</sup></td> | ||
− | <td> | + | <td>150</td> |
− | <td> | + | <td>11</td> |
− | <td></td> | + | <td>1,3x10<sup>7</sup></td> |
− | <td></td> | + | <td>156,6</td> |
</tr> | </tr> | ||
Line 736: | Line 761: | ||
<td>126</td> | <td>126</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>1,3x10<sup>7</sup></td> |
− | <td></td> | + | <td>2,4</td> |
</tr> | </tr> | ||
Line 746: | Line 771: | ||
<td>63</td> | <td>63</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>6,3x10<sup>6</sup></td> |
− | <td></td> | + | <td>1,1%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 753: | Line 778: | ||
<br> | <br> | ||
<h1 class="date three">September 11th</h1> | <h1 class="date three">September 11th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 769: | Line 794: | ||
<td></td> | <td></td> | ||
<td>77</td> | <td>77</td> | ||
− | <td></td> | + | <td>7,7x10<sup>6</sup></td> |
− | <td></td> | + | <td>111,5%</td> |
</tr> | </tr> | ||
Line 779: | Line 804: | ||
<td></td> | <td></td> | ||
<td>149</td> | <td>149</td> | ||
− | <td></td> | + | <td>1,5x10<sup>7</sup></td> |
− | <td></td> | + | <td>176,5</td> |
</tr> | </tr> | ||
Line 788: | Line 813: | ||
<td>5,0x10<sup>8</sup></td> | <td>5,0x10<sup>8</sup></td> | ||
<td>164</td> | <td>164</td> | ||
− | <td></td> | + | <td>/</td> |
− | <td></td> | + | <td>1,6x10<sup>6</sup></td> |
− | <td></td> | + | <td>0,3%</td> |
</tr> | </tr> | ||
Line 799: | Line 824: | ||
<td>299</td> | <td>299</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>3,0x10<sup>6</sup></td> |
− | <td></td> | + | <td>0,5%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 806: | Line 831: | ||
<br> | <br> | ||
<h1 class="date three">September 12th</h1> | <h1 class="date three">September 12th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 822: | Line 847: | ||
<td>877</td> | <td>877</td> | ||
<td>88</td> | <td>88</td> | ||
− | <td></td> | + | <td>1,3x10<sup>7</sup></td> |
− | <td></td> | + | <td>131,3%</td> |
</tr> | </tr> | ||
Line 832: | Line 857: | ||
<td>513</td> | <td>513</td> | ||
<td>52</td> | <td>52</td> | ||
− | <td></td> | + | <td>7,2x10<sup>6</sup></td> |
− | <td></td> | + | <td>109,1%</td> |
</tr> | </tr> | ||
Line 842: | Line 867: | ||
<td>331</td> | <td>331</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>3,3x10<sup>6</sup></td> |
− | <td></td> | + | <td>0,8%</td> |
</tr> | </tr> | ||
Line 852: | Line 877: | ||
<td>368</td> | <td>368</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>3,7x10<sup>6</sup></td> |
− | <td></td> | + | <td>0,8%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 860: | Line 885: | ||
<br> | <br> | ||
<h1 class="date three">September 13th</h1> | <h1 class="date three">September 13th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 876: | Line 901: | ||
<td>/</td> | <td>/</td> | ||
<td>123</td> | <td>123</td> | ||
− | <td></td> | + | <td>1,2x10<sup>7</sup></td> |
− | <td></td> | + | <td>155,8%</td> |
</tr> | </tr> | ||
Line 886: | Line 911: | ||
<td>/</td> | <td>/</td> | ||
<td>117</td> | <td>117</td> | ||
− | <td></td> | + | <td>1,2x10<sup>7</sup></td> |
− | <td></td> | + | <td>134,8%</td> |
</tr> | </tr> | ||
Line 896: | Line 921: | ||
<td>28</td> | <td>28</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>2,8x10<sup>5</sup></td> |
− | <td></td> | + | <td>0,05%</td> |
</tr> | </tr> | ||
Line 906: | Line 931: | ||
<td>30</td> | <td>30</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>3,0x10<sup>5</sup></td> |
− | <td></td> | + | <td>0,07%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 914: | Line 939: | ||
<h1 class="date three">September 14th</h1> | <h1 class="date three">September 14th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 930: | Line 955: | ||
<td>/</td> | <td>/</td> | ||
<td>103</td> | <td>103</td> | ||
− | <td></td> | + | <td>1,0x10<sup>7</sup></td> |
− | <td></td> | + | <td>128,2%</td> |
</tr> | </tr> | ||
Line 940: | Line 965: | ||
<td>/</td> | <td>/</td> | ||
<td>83</td> | <td>83</td> | ||
− | <td></td> | + | <td>8,3x10<sup>6</sup></td> |
− | <td></td> | + | <td>105,1%</td> |
</tr> | </tr> | ||
Line 950: | Line 975: | ||
<td>12</td> | <td>12</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>1,2x10<sup>5</sup></td> |
− | <td></td> | + | <td>0,02%</td> |
</tr> | </tr> | ||
Line 960: | Line 985: | ||
<td>49</td> | <td>49</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>4,9x10<sup>5</sup></td> |
− | <td></td> | + | <td>0,1%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 967: | Line 992: | ||
<br> | <br> | ||
<h1 class="date three">September 15th</h1> | <h1 class="date three">September 15th</h1> | ||
− | <table style="width:100%; text-align:center"> | + | <table style="font-size: 0.7em; width:100%; text-align:center"> |
<tr> | <tr> | ||
<th>Studied strains</th> | <th>Studied strains</th> | ||
Line 981: | Line 1,006: | ||
<td>1,08</td> | <td>1,08</td> | ||
<td>7,1x10<sup>6</sup></td> | <td>7,1x10<sup>6</sup></td> | ||
− | <td></td> | + | <td>/</td> |
− | <td></td> | + | <td>91</td> |
− | <td></td> | + | <td>9,1x10<sup>6</sup></td> |
− | <td></td> | + | <td>128,2</td> |
</tr> | </tr> | ||
Line 991: | Line 1,016: | ||
<td>1,44</td> | <td>1,44</td> | ||
<td>8,5x10<sup>6</sup></td> | <td>8,5x10<sup>6</sup></td> | ||
− | <td></td> | + | <td>/</td> |
− | <td></td> | + | <td>84</td> |
− | <td></td> | + | <td>8,4x10<sup>6</sup></td> |
− | <td></td> | + | <td>98,8</td> |
</tr> | </tr> | ||
Line 1,003: | Line 1,028: | ||
<td>4</td> | <td>4</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>4,0x10<sup>4</sup></td> |
− | <td></td> | + | <td>0,0%</td> |
</tr> | </tr> | ||
Line 1,013: | Line 1,038: | ||
<td>10</td> | <td>10</td> | ||
<td>/</td> | <td>/</td> | ||
− | <td></td> | + | <td>1,0x10<sup>5</sup></td> |
− | <td></td> | + | <td>0,0%</td> |
</tr> | </tr> | ||
</table> | </table> | ||
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Latest revision as of 12:55, 21 October 2015