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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<h2>Goal</h2> | <h2>Goal</h2> | ||
− | + | 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> | |
+ | 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> | ||
+ | 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. | ||
+ | |||
+ | <h2>Material</h2> | ||
+ | <ul> | ||
+ | <li>Glycerol stock of Sc. mcherry</li> | ||
+ | <li>YPD broth</li> | ||
+ | <li>Agar</li> | ||
+ | <li>Geneticin, 100mg/ml</li> | ||
+ | <li>Osmosed water</li> | ||
+ | <li>Sterile culture tubes</li> | ||
+ | <li>Sterile eppendorf tubes</li> | ||
+ | <li>Sterile plates and beads</li> | ||
+ | </ul> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | <img src="https://static.igem.org/mediawiki/2015/5/5f/PBmanufacturingdilutions.jpg" width="400px" style="display:block; margin-right: 30px; float:left;"> | |
− | <img src="https://static.igem.org/mediawiki/2015/5/5f/PBmanufacturingdilutions.jpg" width="400px" | + | <ul><li>Make a culture of Sc. mcherry in 10ml of YPD media adding 20µL of geneticin</li> |
− | + | (We did cultures on the 25th of July) | |
− | + | <li>Two days later, centrifuge the culture tube</li> | |
− | <li> | + | <li>Throw the media away and replace it by 10ml of osmosed water</li> |
− | <li> | + | <li>Measure the OD, doing the blank with osmosed water</li> |
− | <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> | + | <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> |
− | <li> | + | |
− | <li> | + | |
<br> | <br> | ||
− | + | That is what we did today, and we plated the different dilutions, thanks to the beads. | |
− | + | ||
− | + | ||
+ | |||
<br><br> | <br><br> | ||
<h1 class="date one">July 28th</h1> | <h1 class="date one">July 28th</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
− | Our first idea to distribute the yeast | + | Our first idea to distribute the yeast was to simply dry them and collect the dry cells, without adding anything.<br> |
+ | Today is a first try, to have an idea of how to do it.<br> | ||
+ | |||
+ | <h2>Material</h2> | ||
+ | <ul><li>Culture of Sc. mcherry</li> | ||
+ | <li>YPD broth</li> | ||
+ | <li>Agar</li> | ||
+ | <li>Geneticin, 100mg/ml</li> | ||
+ | <li>Osmosed water</li> | ||
+ | <li>Sterile eppendorf tubes</li> | ||
+ | <li>Sterile plates and beads</li></ul> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
<ul> | <ul> | ||
− | <li>Centrifuge | + | <li>Centrifuge a culture tube</li> |
<li>Throw the media away</li> | <li>Throw the media away</li> | ||
− | <li>Add | + | <li>Add 300µL of water to detach the yeasts from the tube easily</li> |
<li>Spread on aluminium</li> | <li>Spread on aluminium</li> | ||
− | <li>Let it dry for 2 hours</li> | + | <li>Let it dry for 2 hours in the open air or in the incubator (2 different tests)</li> |
<li>Scratch the aluminium to detach the dried yeasts</li> | <li>Scratch the aluminium to detach the dried yeasts</li> | ||
− | <li> | + | <li>Put the powder in 10ml of osmosed water</li> |
+ | <li>Plate the solution to see if Sc. mcherry survived</li> | ||
</ul> | </ul> | ||
− | <h2>Results</h2> | + | |
+ | <h2>Results and discussion</h2> | ||
<div class="column-left"><p> | <div class="column-left"><p> | ||
<br> | <br> | ||
We obtained this powder:<br><br> | We obtained this powder:<br><br> | ||
− | + | We realised the aluminium was not a good idea:<br> | |
The yeasts stick to it and it gets destroyed when we scratch.<br> | The yeasts stick to it and it gets destroyed when we scratch.<br> | ||
We have to find another drying paper, more resistant.<br> | We have to find another drying paper, more resistant.<br> | ||
Line 61: | Line 89: | ||
</p></div> | </p></div> | ||
<div style="clear: both"></div> | <div style="clear: both"></div> | ||
+ | |||
<br> | <br> | ||
<h1 class="date one">July 29th</h1> | <h1 class="date one">July 29th</h1> | ||
− | <H2>Result of the | + | |
− | The 10ml solution of | + | <H2>Result of the OD test made on the 27th of July:</h2> |
− | <b>1 | + | The 10ml solution of Sc. mcherry/water had an OD of 2,557.<br> |
− | We | + | 100µL of the 10<sup>-4</sup> dilution grew 359 colonies, so:<br> |
+ | <b>1 A = 1,4x10<sup>7</sup> cells/ml</b><br> | ||
+ | We repeated the very same OD experiment today, for more repetitions. | ||
+ | |||
<br><br> | <br><br> | ||
<h1 class="date one">July 30th</h1> | <h1 class="date one">July 30th</h1> | ||
− | |||
− | |||
− | |||
− | |||
− | <br><br> | + | <h2>Results of the drying test made on the 28th of July:</h2> |
+ | Yeasts grew in the plate.<br> | ||
+ | We can check that they produce RFP, it's Sc. mcherry who survived.<br> | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | 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<sup>-1</sup> to 10<sup>-5</sup>)</li> | ||
+ | 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>Centrifuge the culture tube</li> | ||
+ | <li>Throw the media away</li> | ||
+ | <li>Add 300µL of water to detach the yeasts from the tube easily</li> | ||
+ | <li>Spread on aluminium</li> | ||
+ | <li>Let it dry for 2 hours</li> | ||
+ | <li>Scratch the aluminium to detach the dried yeasts</li> | ||
+ | <li>Put the powder in 10ml of osmosed water</li> | ||
+ | <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> | ||
+ | |||
+ | |||
+ | <br> | ||
<h1 class="date one">July 31th</h1> | <h1 class="date one">July 31th</h1> | ||
− | <h2>Result of the | + | <h2>Result of the OD test made on the 29th of July:</h2> |
− | <b>1 | + | 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> | ||
+ | <b>1 A = 7,8x10<sup>6</sup> cells/ml</b> | ||
− | <br>< | + | |
+ | <br> | ||
+ | <a name="august" class="anchor"><h1></h1></a> | ||
<h1 class="date two">August 1st</h1> | <h1 class="date two">August 1st</h1> | ||
− | <h2>Goal</h2> | + | <h2>Result of the drying test made on the 30th of July:</h2> |
− | The powder obtained with the drying method | + | 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<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%. | ||
+ | |||
+ | <h2>Discussion and New Goal</h2> | ||
+ | The powder we obtained with the drying method stick to everything, because of static electricity and therefore, it is not very easy to distribute and manipulate.<br> | ||
+ | Consequently, we are looking fo other solutions...<br> | ||
<br> | <br> | ||
− | Our second idea | + | 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="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> | |
<ul> | <ul> | ||
− | <li> | + | <li>Test 1, original recipe: wheat flour, sugar and ginger </li> |
− | <li> | + | Since the project is designed for rice fermentation and rice is cheaper than wheat, we wanted to try with rice flour. |
− | <li> | + | <li>Test 2: rice flour, sugar and ginger</li> |
− | <li> | + | Ginger can be expensive so maybe we can avoid using it... |
+ | <li>Test 3: rice flour, sugar</li> | ||
+ | And maybe we can also avoid using sugar | ||
+ | <li>Test 4: only rice flour</li> | ||
</ul> | </ul> | ||
+ | |||
+ | <h2>Material</h2> | ||
+ | <ul> | ||
+ | <li>Culture of Sc. mcherry</li> | ||
+ | <li>Potatoes</li> | ||
+ | <li>Flour (wheat and rice)</li> | ||
+ | <li>Cornmeal</li> | ||
+ | <li>Sugar</li> | ||
+ | <li>Ginger</li> | ||
+ | <li>Plate of YPDagar+Gen(200)</li> | ||
+ | <li>Osmosed water</li> | ||
+ | <li>Sterile eppendorf tubes</li> | ||
+ | <li>Sterile beads</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Procedure</h2> | ||
<div class="column-left"> | <div class="column-left"> | ||
<ul> | <ul> | ||
+ | <li>Cook some potatoes</li> | ||
+ | <li>Save the cooking water, wait till it's not too hot and mix 20ml of it with the yeasts</li> | ||
+ | <li>Add 20ml of flour (rice or wheat depending on the recipe), 40ml of mashed potatoes</li> | ||
+ | <li>Depending on the recipe add 20ml of sugar and a teaspoon of ginger</li> | ||
+ | <li>Add 80ml of cornmeal</li> | ||
<li>Let it dry for 1 day</li> | <li>Let it dry for 1 day</li> | ||
<li>Mill the paste to obtain a powder</li> | <li>Mill the paste to obtain a powder</li> | ||
− | <li>Plate the powder obtained to see how many Sc. | + | <li>Plate the powder obtained to see how many Sc. mcherry survived </li> |
</ul><br> | </ul><br> | ||
− | |||
− | |||
</div> | </div> | ||
<div class="column-right"><p> | <div class="column-right"><p> | ||
Line 112: | Line 191: | ||
<br> | <br> | ||
<h1 class="date two">August 3rd</h1> | <h1 class="date two">August 3rd</h1> | ||
− | <h2> | + | <h2>Continuing the powder recipe we began on the 1st of August</h2> |
− | + | ||
− | + | ||
<div class="column-left"><p> | <div class="column-left"><p> | ||
<br> | <br> | ||
− | + | After almost two days of drying, we obtained a very hard paste that we milled.<br> | |
− | We put | + | After milling, we had this nice yellow powder you can see on the picture.<br><br> |
− | + | We put 100mg of each powder in 10ml of osmosed water, made dilutions,<br> | |
+ | and plated the solutions in YPDagar+GEN(200) plates. | ||
</p></div> | </p></div> | ||
<div class="column-right"><p> | <div class="column-right"><p> | ||
Line 125: | Line 203: | ||
</p></div> | </p></div> | ||
<div style="clear: both"></div> | <div style="clear: both"></div> | ||
+ | |||
<br> | <br> | ||
<h1 class="date two">August 4th</h1> | <h1 class="date two">August 4th</h1> | ||
− | <h2>Goal</h2> | + | <h2>Discussion and New Goal</h2> |
− | While doing the recipe with the potatoes, we | + | While doing the recipe with the potatoes (from the 1st to the 3rd of August), we realised it wasn't easy to do and it took lots of time and ingredients.<br> |
We decided to reduce the number of ingredients, using only water and rice flour.<br> | We decided to reduce the number of ingredients, using only water and rice flour.<br> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
<ul> | <ul> | ||
− | <li>Mix the yeasts with | + | <li>Mix the yeasts with 20ml of osmosed water</li> |
− | <li>Add | + | <li>Add 40ml of rice flour</li> |
<li>Knead till you get a nice paste</li> | <li>Knead till you get a nice paste</li> | ||
</ul> | </ul> | ||
Line 148: | Line 228: | ||
Several cubes can be packed together.<br><br> | Several cubes can be packed together.<br><br> | ||
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 | + | We dissolved one cube in 10ml of osmosed water, after weighed it.<br> |
+ | 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> | ||
Line 154: | Line 235: | ||
</p></div> | </p></div> | ||
<div style="clear: both"></div> | <div style="clear: both"></div> | ||
+ | |||
<br> | <br> | ||
<h1 class="date two">August 5th</h1> | <h1 class="date two">August 5th</h1> | ||
− | <h2>Results of the potato powder:</h2> | + | <h2>Results of the survival of Sc. mcherry in the potato powder made on the 3rd of August:</h2> |
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> | ||
− | We can see that the more ingredients we put, the more the yeasts survived. | + | <h2>Discussion:</h2> |
+ | We can see that the more ingredients we put, the more the yeasts survived, so each ingredient plays a role in the cells survival.<br> | ||
+ | But as we said before, we don't want to put too much ingredients in the recipe so that it's cheap and easy.<br> | ||
+ | We can also notice that rice flour seems more efficient than wheat flour (but we made only one repetition so nothing is statistically proven). | ||
<br><br> | <br><br> | ||
<h1 class="date two">August 6th</h1> | <h1 class="date two">August 6th</h1> | ||
− | <h2>Results of the | + | <h2>Results of the survival of Sc. mcherry in the cubes made on the 4th of August:</h2> |
− | + | <div class="column-left"><p> | |
+ | <br> | ||
+ | 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> | ||
+ | <div class="column-right"><p> | ||
+ | <img src="https://static.igem.org/mediawiki/2015/8/85/PBmanufacturingwaterbath.jpg" width="350px"><br> | ||
+ | </p></div> | ||
+ | <div style="clear: both"></div> | ||
− | + | <br> | |
<h1 class="date two">August 7th</h1> | <h1 class="date two">August 7th</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
Now that we are at ease with the process, we are trying the very same process with lactococcus lactis. | Now that we are at ease with the process, we are trying the very same process with lactococcus lactis. | ||
− | We chose MG 13.63, containing the plasmid PSIP411, resistant to erythromycin at a concentration of 10µL/mL, to study it's survival rate. We will name this strain G1513. We also have to correlate the OD of a solution of G1513 to the number of alive cells inside. | + | We chose MG 13.63, containing the plasmid PSIP411, resistant to erythromycin at a concentration of 10µL/mL, to study it's survival rate.<br> |
+ | We will name this strain G1513.<br> | ||
+ | We also have to correlate the OD of a solution of G1513 to the number of alive cells inside. | ||
− | < | + | <h2>Material</h2> |
+ | <ul> | ||
+ | <li>Glycerol stock of G1513</li> | ||
+ | <li>M17 broth</li> | ||
+ | <li>Solution of D-Glucose</li> | ||
+ | <li>Agar</li> | ||
+ | <li>Erythromycin, 20mg/ml</li> | ||
+ | <li>Osmosed water</li> | ||
+ | <li>Sterile eppendorf tubes</li> | ||
+ | <li>Sterile plates and beads</li> | ||
+ | </ul> | ||
+ | |||
+ | <h2>Procedure</h2> | ||
+ | <ul><li>Make a culture of G1513 in 10ml of M17+1% glucose media adding 5µL of erythromycin</li> | ||
+ | (We did cultures on the 6th of August) | ||
+ | <li>One day later, centrifuge the culture tube</li> | ||
+ | <li>Throw the media away and replace it by 10ml of 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>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> | ||
<h1 class="date two">August 10th</h1> | <h1 class="date two">August 10th</h1> | ||
− | <h2>Results of the OD test for G1513:</h2> | + | <h2>Results of the OD test for G1513 made on the 7th of August:</h2> |
− | <b>1 | + | The 10ml solution of G1513/water had an OD of 1,296.<br> |
+ | Here is a table of what 100µL of each dilutions grew: | ||
+ | <table style="width:25%" align="center"> | ||
+ | <tr style="background-color:#E6E6E6"> | ||
+ | <th>Dilution </th> | ||
+ | <th>Number of colonies</th> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-1</sup></td> | ||
+ | <td>381</td> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-2</sup></td> | ||
+ | <td>84</td> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-3</sup></td> | ||
+ | <td>11</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | We have, in average, 7,74x10<sup>3</sup> cells per ml of the 10<sup>-1</sup> dilution.<br> | ||
+ | so: <b>1 A = 6,0x10<sup>4</sup> cells/ml</b><br> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | Today, we made new cubes containing Sc. mcherry and G1513 to calculate the survival rate. | |
<br><br> | <br><br> | ||
<h1 class="date two">August 11th</h1> | <h1 class="date two">August 11th</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
− | While waiting for the cubes results, we are thinking about the next step: finding a media to grow the strains, easy to give to the population and to use afterward. The perfect media would be an edible one, that we could incorporate to the cube, instead of the water.<br> | + | While waiting for the cubes results, we are thinking about the next step: finding a media to grow the strains, easy to give to the population and to use afterward.<br> |
+ | The perfect media would be an edible one, that we could incorporate to the cube, instead of the water.<br> | ||
+ | |||
+ | <h2>Material</h2> | ||
+ | <ul><li>Culture of Sc. mcherry</li> | ||
+ | <li>3 potatoes</li> | ||
+ | <li>Rice flour</li> | ||
+ | <li>Geneticin, 100mg/ml</li> | ||
+ | <li>Osmosed water</li> | ||
+ | <li>Sterile eppendorf tubes and culture tubes</li> | ||
+ | <li>Sterile plates and beads</li> | ||
+ | <li>One glass bottle</li></ul> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
We tried 2 different medium: potato juice and potato juice with rice flour.<br><br> | We tried 2 different medium: potato juice and potato juice with rice flour.<br><br> | ||
Line 197: | Line 347: | ||
<li>Let the water boil (sterilisation)</li> | <li>Let the water boil (sterilisation)</li> | ||
<li>Save the water and stock it in a bottle </li> | <li>Save the water and stock it in a bottle </li> | ||
− | <li>Put 10ml of this | + | <li>Put 10ml of this potato water in a culture tube</li> |
<li>Add 20μL of geneticin (100mg/ml)</li> | <li>Add 20μL of geneticin (100mg/ml)</li> | ||
− | <li>Inoculate with Sc. | + | <li>Inoculate with Sc. mcherry</li> |
<li>Put in the incubator at 30°C for 2 days</li> | <li>Put in the incubator at 30°C for 2 days</li> | ||
</ul><br> | </ul><br> | ||
For the mix potato juice with rice flour:<br> | For the mix potato juice with rice flour:<br> | ||
<ul> | <ul> | ||
− | <li>Put 5ml of the | + | <li>Put 5ml of the potato water made previously in a culture tube</li> |
− | <li>Inoculate with Sc. | + | <li>Inoculate with Sc. mcherry</li> |
<li>Add 20μL of geneticin (100mg/ml)</li> | <li>Add 20μL of geneticin (100mg/ml)</li> | ||
<li>Add 5ml of rice flour</li> | <li>Add 5ml of rice flour</li> | ||
Line 213: | Line 363: | ||
<br> | <br> | ||
<h1 class="date two">August 12th</h1> | <h1 class="date two">August 12th</h1> | ||
− | <h2>Results of the cube tests</h2> | + | <h2>Results of the cube tests made on the 10th of August</h2> |
The yeasts survived well: 42%. <br> | The yeasts survived well: 42%. <br> | ||
− | Seven days later, we plated | + | (Seven days later, on the 19th of August, we plated cubes from this same test, and obtained a survival rate of 21%.)<br> |
− | But strangely, the number of cells in the plates of | + | But about the G1513, strangely, the number of cells in the plates of M17agar, which are supposed to be G1513, is the same as in the plates with YPD and geneticin, selecting the yeasts.<br> |
− | Therefore, we checked the production of RFP, since Sc. | + | Therefore, we checked the production of RFP, since Sc. mcherry produces RFP but not G1513.<br> |
− | Unfortunately, we came to the conclusion that the cells in the M17 plates were yeasts. Actually, Sc. | + | Unfortunately, we came to the conclusion that the cells in the M17 plates were yeasts. Actually, Sc. mcherry resist to erythromycin and is therefore growing in the plate.<br> |
+ | |||
<h2>Goal</h2> | <h2>Goal</h2> | ||
− | We need a chemical that will allow us to select G1513 and kill Sc. | + | We need a chemical that will allow us to select G1513 and kill Sc. mcherry.<br> |
After some researchs, we found out that cycloheximide could be helpful and ordered it. While waiting for this chemical, we can't work with G1513. | After some researchs, we found out that cycloheximide could be helpful and ordered it. While waiting for this chemical, we can't work with G1513. | ||
<br><br> | <br><br> | ||
<h1 class="date two">August 13th</h1> | <h1 class="date two">August 13th</h1> | ||
− | <h2>Results of the media test</h2> | + | <h2>Results of the media test made on the 11th of August</h2> |
Nothing grew on the potato juice media.<br> | Nothing grew on the potato juice media.<br> | ||
We had the idea of adding sugar, to see if it changes something.<br> | We had the idea of adding sugar, to see if it changes something.<br> | ||
Line 233: | Line 384: | ||
<h1 class="date two">August 17th</h1> | <h1 class="date two">August 17th</h1> | ||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | Making new cubes with Sc. | + | Making new cubes with Sc. mcherry alone, G1513 alone, and a mix of both strains. <br> |
Measuring the OD and plating to count the cells | Measuring the OD and plating to count the cells | ||
− | <h2>Results of the media test</h2> | + | <h2>Results of the media test made on the 13th of August</h2> |
With sugar, we can see there are cells growing in the potato juice.<br> | With sugar, we can see there are cells growing in the potato juice.<br> | ||
It's a very small growth compared to the one in YPD but it's still a growth.<br> | It's a very small growth compared to the one in YPD but it's still a growth.<br> | ||
− | We will work more precisely on this media, trying to deduce the influence of the | + | We will work more precisely on this media, trying to deduce the influence of the potato and sugar concentration. |
<br><br> | <br><br> | ||
<h1 class="date two">August 18th</h1> | <h1 class="date two">August 18th</h1> | ||
− | <h2>Results of the OD test</h2> | + | <h2>Results of the OD test made on the 17th of August</h2> |
− | For Sc. | + | For Sc. mcherry:<br> |
− | <b>1 | + | 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> | ||
+ | <b>1 A = 3,8x10<sup>6</sup> cells/ml</b><br> | ||
+ | |||
For G1513:<br> | For G1513:<br> | ||
− | <b>1 | + | The 10ml solution of G1513/water had an OD of 1,521.<br> |
+ | 100µL of the dilutions grew:<br> | ||
+ | <table style="width:25%" align="center"> | ||
+ | <tr style="background-color:#E6E6E6"> | ||
+ | <th>Dilution </th> | ||
+ | <th>Number of colonies</th> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-5</sup></td> | ||
+ | <td>131</td> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-6</sup></td> | ||
+ | <td>21</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | So <b>1 A = 1,1x10<sup>8</sup> cells/ml</b> | ||
+ | |||
<br><br> | <br><br> | ||
<h1 class="date two">August 19th</h1> | <h1 class="date two">August 19th</h1> | ||
− | <h2>Results of the cube test</h2> | + | <h2>Results of the cube test made on the 17th of August</h2> |
<ul> | <ul> | ||
− | <li>Sc. | + | <li>Sc. mcherry alone: 39%</li> |
− | <li>Sc. | + | <li>Sc. mcherry in the mix: 26%</li> |
+ | <li>G1513 in the mix: not plated because we are waiting for the cycloheximide</li> | ||
<li>G1513 alone: 1,8%</li> | <li>G1513 alone: 1,8%</li> | ||
</ul> | </ul> | ||
We kept samples of the mix in the fridge to be able to study it later, when we will have the cycloheximide. | We kept samples of the mix in the fridge to be able to study it later, when we will have the cycloheximide. | ||
+ | |||
<br><br> | <br><br> | ||
<h1 class="date two">August 20th</h1> | <h1 class="date two">August 20th</h1> | ||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | We made more cubes with Sc. mcherry alone, G1513 alone, and a mix of both.<br> | |
− | + | We made more OD correlations. | |
+ | |||
<br><br> | <br><br> | ||
<h1 class="date two">August 24th</h1> | <h1 class="date two">August 24th</h1> | ||
+ | |||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | We made more cubes...<br> | |
− | <h2>Results of the OD test</h2> | + | <h2>Results of the OD test made on the 20th of August</h2> |
− | For Sc. | + | For Sc. mcherry:<br> |
− | <b>1 | + | 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> | ||
+ | <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> | ||
− | <h2>Results of the cube test</h2> | + | <h2>Results of the cube test made on the 20th of August</h2> |
<ul> | <ul> | ||
− | <li>Sc. | + | <li>Sc. mcherry alone: 92%</li> |
− | <li>Sc. | + | <li>Sc. mcherry in the mix (but dead G1513): 96%</li> |
<li>G1513 alone: Nothing</li> | <li>G1513 alone: Nothing</li> | ||
</ul> | </ul> | ||
Line 284: | Line 461: | ||
<br> | <br> | ||
<h1 class="date two">August 25th</h1> | <h1 class="date two">August 25th</h1> | ||
− | <h2>Results of the OD | + | <h2>Results of the OD tests made on the 24th of August</h2> |
− | For Sc. | + | For Sc. mcherry:<br> |
− | <b>1 | + | The first 10ml solution of Sc. mcherry/water had an OD of 1,921.<br> |
+ | 100µL of the dilutions grew:<br> | ||
+ | <table style="width:25%" align="center"> | ||
+ | <tr style="background-color:#E6E6E6"> | ||
+ | <th>Dilution </th> | ||
+ | <th>Number of colonies</th> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-4</sup></td> | ||
+ | <td>505</td> | ||
+ | </tr> | ||
+ | <tr align="center"> | ||
+ | <td>10<sup>-5</sup></td> | ||
+ | <td>39</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <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> | ||
+ | 100µL of the 10<sup>-5</sup> dilution grew 95 colonies.<br> | ||
+ | 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. | ||
+ | |||
<br><br> | <br><br> | ||
<h1 class="date two">August 26th</h1> | <h1 class="date two">August 26th</h1> | ||
− | <p><img src="https://static.igem.org/mediawiki/2015/d/d9/PBmanufacturingcubeenmain.jpg" width=" | + | <h2>Results of the cube test made on the 24th of August</h2> |
− | + | <p><img src="https://static.igem.org/mediawiki/2015/d/d9/PBmanufacturingcubeenmain.jpg" width="150" style="display:block; margin-right: 20px; float:left;"> | |
− | <li>Sc. | + | <li>Sc. mcherry alone: 122% et 162%</li><br> |
− | <li>Sc. | + | <li>Sc. mcherry in the mix (but dead G1513): 178% et 217%</li><br> |
<li>G1513 alone: Nothing</li> | <li>G1513 alone: Nothing</li> | ||
</p> | </p> | ||
− | + | <br> | |
<h1 class="date two">August 27th</h1> | <h1 class="date two">August 27th</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
We want to find a media that could be use in India to grow the yeasts and the Lactococcus lactis for cheap.<br> | We want to find a media that could be use in India to grow the yeasts and the Lactococcus lactis for cheap.<br> | ||
+ | <h2>Material</h2> | ||
+ | <ul> | ||
+ | <li>Potatoes</li> | ||
+ | <li>Sugar</li> | ||
+ | <li>Water</li> | ||
+ | <li>Culture of Sc. mcherry</li> | ||
+ | <li>Culture of G1513</li> | ||
+ | <li>Erythromycin, Geneticin</li> | ||
+ | <li>Tecan and Tecan plate</li> | ||
+ | <li>Mineral oil</li> | ||
+ | </ul> | ||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
<ul> | <ul> | ||
− | <li>Make 3 different potato juice solutions | + | <li>Make 3 different potato juice solutions(low, medium and high concentration of potatoes) following this protocol</li> |
− | <li>For each one of these solutions, make samples with different concentration of sugar</li> | + | <li>For each one of these solutions, make samples with different concentration of sugar: 0g/ml; 0,1g/ml; 0,2g/ml; 0,4g/ml.</li> |
− | <li>Inoculate the strains | + | <li>Inoculate the strains in these media, with the appropriate antibiotics (do not mix G1513 and Sc. mcherry)</li> |
− | <li>Put 150μL of the solutions in | + | <li>Put 150μL of the solutions in Tecan plate wells and add 50μL of oil</li> |
<li>Put in the Tecan at 30°C, measuring the OD every 15minutes, for 2 days</li> | <li>Put in the Tecan at 30°C, measuring the OD every 15minutes, for 2 days</li> | ||
</ul> | </ul> | ||
Line 315: | Line 523: | ||
<h1 class="date two">August 28th</h1> | <h1 class="date two">August 28th</h1> | ||
<h2>Goal</h2> | <h2>Goal</h2> | ||
− | We want to see how the strains in the cubes react when put in | + | We want to see how the strains in the cubes react when put in idli.<br> |
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | <ul> | |
+ | <li>Following the <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#FSIdli" target="_blank">idli recipe protocol</a>, make idli.</li> | ||
+ | <li>Weigh the cube</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>After the fermentation, plate 100μL of a 10<sup>-2</sup> dilution of the Idli water</li> | ||
+ | </ul> | ||
− | + | ||
+ | <br> | ||
<h1 class="date two">August 31th</h1> | <h1 class="date two">August 31th</h1> | ||
− | <h2>Results of the media test</h2> | + | <h2>Results of the media test made on the 27th of August</h2> |
It's a fail, the OD was too high so we can't really see the growth.<br> | It's a fail, the OD was too high so we can't really see the growth.<br> | ||
− | We did it again, diluting the media | + | We did it again, diluting the inoculated media 200times in osmosed water.<br> |
+ | |||
<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"><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</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> | ||
− | We will do it again, asking for a real protocol to | + | We will do it again, asking for a real protocol to our advisors, because obviously, there was a misunderstanding. |
+ | |||
<br><br> | <br><br> | ||
<h1 class="date three">September 4th</h1> | <h1 class="date three">September 4th</h1> | ||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | We tried the new method for the Tecan test.<br> | |
This time, we did an overnight culture for each media and each strain, that we diluted 200times (till we get an OD smaller than 0,01) | This time, we did an overnight culture for each media and each strain, that we diluted 200times (till we get an OD smaller than 0,01) | ||
+ | |||
+ | |||
+ | <br><br> | ||
+ | <h1 class="date three">September 6th</h1> | ||
+ | <h2>Procedure</h2> | ||
+ | We made fresh new cultures of G1513 and Sc. mcherry. | ||
<br><br> | <br><br> | ||
<h1 class="date three">September 7th</h1> | <h1 class="date three">September 7th</h1> | ||
− | <h2>Results of the Tecan test</h2> | + | <h2>Results of the Tecan test made on the 4th of September</h2> |
Third fail, because one parameter was changed and we didn't noticed: the Tecan measured the OD for only few hours, not enough to see the growth... | Third fail, because one parameter was changed and we didn't noticed: the Tecan measured the OD for only few hours, not enough to see the growth... | ||
Line 347: | Line 589: | ||
<h1 class="date three">September 8th</h1> | <h1 class="date three">September 8th</h1> | ||
<h2>Procedure</h2> | <h2>Procedure</h2> | ||
− | + | Today we used the fresh cultures we made on the 6th of August to make lots of new cubes and OD test.<br> | |
− | + | 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> | ||
+ | <li>Solution Y: 5,2x10<sup>9</sup> cells of G1513 per ml</li></ul> | ||
− | <br><br> | + | We made 3 different types of cubes: |
+ | <ul> | ||
+ | <li>Cubes Q: 15ml of rice flour, 6ml of osmosed water and 1ml of solution X</li> | ||
+ | <li>Cubes R: 15ml of rice flour, 5ml of osmosed water, 900μL of solution X and 1ml of solution Y</li> | ||
+ | <li>Cubes S: 15ml of rice flour, 6ml of osmosed water and 1ml of solution Y</li> | ||
+ | </ul> | ||
+ | <table style="width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Strains and cubes</th> | ||
+ | <th>Total weight of the cubes (g)</th> | ||
+ | <th>Number of cells per g of cube</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q cubes</td> | ||
+ | <td>13,30</td> | ||
+ | <td>6,6x10<sup>6</sup></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in R cubes</td> | ||
+ | <td>13,39</td> | ||
+ | <td>5,9x10<sup>6</sup></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>G1513 in R cubes</td> | ||
+ | <td>13,39</td> | ||
+ | <td>3,9x10<sup>8</sup></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in S cubes</td> | ||
+ | <td>10,70</td> | ||
+ | <td>4,8x10<sup>8</sup></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | We are going to plate one cube of each kind each day and follow the survival rate during a week of drying. | ||
+ | |||
+ | For the OD, here are the results we obtained (we obtained them on the 10th of August). | ||
+ | <table style="width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>OD of the sample</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution of the sample</th> | ||
+ | <th>Number of colonies in the 10<sup>-4</sup> dilution of the sample</th> | ||
+ | <th>Number of colonies in the 10<sup>-5</sup> dilution of the sample</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry</td> | ||
+ | <td>1,156</td> | ||
+ | <td>+++</td> | ||
+ | <td>347</td> | ||
+ | <td>23</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513</td> | ||
+ | <td>0,392</td> | ||
+ | <td>+++</td> | ||
+ | <td>439</td> | ||
+ | <td>/</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | So, for Sc. mcherry:<b> 1A=2,5x10<sup>7</sup> cells/ml </b> <br> | ||
+ | For G1513:<b> 1A=1,1x10<sup>8</sup> cells/ml </b> | ||
+ | |||
+ | |||
+ | <br> | ||
<h1 class="date three">September 9th</h1> | <h1 class="date three">September 9th</h1> | ||
+ | We plated the first cubes today.<br> | ||
+ | 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). | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-4</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>0,96</td> | ||
+ | <td>6,3x10<sup>6</sup></td> | ||
+ | <td>72</td> | ||
+ | <td>10</td> | ||
+ | <td>8,6x10<sup>6</sup></td> | ||
+ | <td>136,5%</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>0,96</td> | ||
+ | <td>5,7x10<sup>6</sup></td> | ||
+ | <td>80</td> | ||
+ | <td>11</td> | ||
+ | <td>9,5x10<sup>6</sup></td> | ||
+ | <td>166,7%</td> | ||
+ | </tr> | ||
− | < | + | <tr> |
+ | <td>G1513 in R</td> | ||
+ | <td>0,96</td> | ||
+ | <td>3,7x10<sup>8</sup></td> | ||
+ | <td>79</td> | ||
+ | <td>10</td> | ||
+ | <td>9,0x10<sup>6</sup></td> | ||
+ | <td>2,4%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>0,72</td> | ||
+ | <td>5,4x10<sup>8</sup></td> | ||
+ | <td>357</td> | ||
+ | <td>23</td> | ||
+ | <td>2,9x10<sup>7</sup></td> | ||
+ | <td>5,4%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
<h1 class="date three">September 10th</h1> | <h1 class="date three">September 10th</h1> | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-4</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>0,97</td> | ||
+ | <td>6,4x10<sup>6</sup></td> | ||
+ | <td>104</td> | ||
+ | <td>5</td> | ||
+ | <td>7,7x10<sup>6</sup></td> | ||
+ | <td>120,3</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,40</td> | ||
+ | <td>8,3x10<sup>6</sup></td> | ||
+ | <td>150</td> | ||
+ | <td>11</td> | ||
+ | <td>1,3x10<sup>7</sup></td> | ||
+ | <td>156,6</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,40</td> | ||
+ | <td>5,5x10<sup>8</sup></td> | ||
+ | <td>126</td> | ||
+ | <td>/</td> | ||
+ | <td>1,3x10<sup>7</sup></td> | ||
+ | <td>2,4</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>1,19</td> | ||
+ | <td>5,7x10<sup>8</sup></td> | ||
+ | <td>63</td> | ||
+ | <td>/</td> | ||
+ | <td>6,3x10<sup>6</sup></td> | ||
+ | <td>1,1%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
+ | <h1 class="date three">September 11th</h1> | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>1,04</td> | ||
+ | <td>6,9x10<sup>6</sup></td> | ||
+ | <td></td> | ||
+ | <td>77</td> | ||
+ | <td>7,7x10<sup>6</sup></td> | ||
+ | <td>111,5%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,28</td> | ||
+ | <td>8,5x10<sup>6</sup></td> | ||
+ | <td></td> | ||
+ | <td>149</td> | ||
+ | <td>1,5x10<sup>7</sup></td> | ||
+ | <td>176,5</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,28</td> | ||
+ | <td>5,0x10<sup>8</sup></td> | ||
+ | <td>164</td> | ||
+ | <td>/</td> | ||
+ | <td>1,6x10<sup>6</sup></td> | ||
+ | <td>0,3%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>1,49</td> | ||
+ | <td>5,8x10<sup>8</sup></td> | ||
+ | <td>299</td> | ||
+ | <td>/</td> | ||
+ | <td>3,0x10<sup>6</sup></td> | ||
+ | <td>0,5%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
+ | <h1 class="date three">September 12th</h1> | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>1,50</td> | ||
+ | <td>9,9x10<sup>6</sup></td> | ||
+ | <td>877</td> | ||
+ | <td>88</td> | ||
+ | <td>1,3x10<sup>7</sup></td> | ||
+ | <td>131,3%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,12</td> | ||
+ | <td>6,6x10<sup>6</sup></td> | ||
+ | <td>513</td> | ||
+ | <td>52</td> | ||
+ | <td>7,2x10<sup>6</sup></td> | ||
+ | <td>109,1%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,12</td> | ||
+ | <td>4,4x10<sup>8</sup></td> | ||
+ | <td>331</td> | ||
+ | <td>/</td> | ||
+ | <td>3,3x10<sup>6</sup></td> | ||
+ | <td>0,8%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>0,93</td> | ||
+ | <td>4,5x10<sup>8</sup></td> | ||
+ | <td>368</td> | ||
+ | <td>/</td> | ||
+ | <td>3,7x10<sup>6</sup></td> | ||
+ | <td>0,8%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <br> | ||
+ | <h1 class="date three">September 13th</h1> | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>1,17</td> | ||
+ | <td>7,7x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>123</td> | ||
+ | <td>1,2x10<sup>7</sup></td> | ||
+ | <td>155,8%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,50</td> | ||
+ | <td>8,9x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>117</td> | ||
+ | <td>1,2x10<sup>7</sup></td> | ||
+ | <td>134,8%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,50</td> | ||
+ | <td>5,9x10<sup>8</sup></td> | ||
+ | <td>28</td> | ||
+ | <td>/</td> | ||
+ | <td>2,8x10<sup>5</sup></td> | ||
+ | <td>0,05%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>0,96</td> | ||
+ | <td>4,6x10<sup>8</sup></td> | ||
+ | <td>30</td> | ||
+ | <td>/</td> | ||
+ | <td>3,0x10<sup>5</sup></td> | ||
+ | <td>0,07%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
+ | <h1 class="date three">September 14th</h1> | ||
+ | |||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>1,18</td> | ||
+ | <td>7,8x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>103</td> | ||
+ | <td>1,0x10<sup>7</sup></td> | ||
+ | <td>128,2%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,33</td> | ||
+ | <td>7,9x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>83</td> | ||
+ | <td>8,3x10<sup>6</sup></td> | ||
+ | <td>105,1%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,33</td> | ||
+ | <td>5,2x10<sup>8</sup></td> | ||
+ | <td>12</td> | ||
+ | <td>/</td> | ||
+ | <td>1,2x10<sup>5</sup></td> | ||
+ | <td>0,02%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>0,82</td> | ||
+ | <td>3,9x10<sup>8</sup></td> | ||
+ | <td>49</td> | ||
+ | <td>/</td> | ||
+ | <td>4,9x10<sup>5</sup></td> | ||
+ | <td>0,1%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
+ | <h1 class="date three">September 15th</h1> | ||
+ | <table style="font-size: 0.7em; width:100%; text-align:center"> | ||
+ | <tr> | ||
+ | <th>Studied strains</th> | ||
+ | <th>Weight (g)</th> | ||
+ | <th>Number of cells initially in the cube</th> | ||
+ | <th>Number of colonies in the 10<sup>-2</sup> dilution</th> | ||
+ | <th>Number of colonies in the 10<sup>-3</sup> dilution</th> | ||
+ | <th>Number of cells in the cube after the drying period</th> | ||
+ | <th>Survival rate</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sc. mcherry in Q</td> | ||
+ | <td>1,08</td> | ||
+ | <td>7,1x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>91</td> | ||
+ | <td>9,1x10<sup>6</sup></td> | ||
+ | <td>128,2</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>Sc. mcherry in R</td> | ||
+ | <td>1,44</td> | ||
+ | <td>8,5x10<sup>6</sup></td> | ||
+ | <td>/</td> | ||
+ | <td>84</td> | ||
+ | <td>8,4x10<sup>6</sup></td> | ||
+ | <td>98,8</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in R</td> | ||
+ | <td>1,44</td> | ||
+ | <td>5,6x10<sup>8</sup></td> | ||
+ | <td>4</td> | ||
+ | <td>/</td> | ||
+ | <td>4,0x10<sup>4</sup></td> | ||
+ | <td>0,0%</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td>G1513 in S</td> | ||
+ | <td>1,01</td> | ||
+ | <td>4,9x10<sup>8</sup></td> | ||
+ | <td>10</td> | ||
+ | <td>/</td> | ||
+ | <td>1,0x10<sup>5</sup></td> | ||
+ | <td>0,0%</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
</html> | </html> |
Latest revision as of 12:55, 21 October 2015