Difference between revisions of "Team:Paris Bettencourt/Notebook/Manufacturing"

Line 7: Line 7:
 
Beginning of the manufacturing project! Let's do it!<br>
 
Beginning of the manufacturing project! Let's do it!<br>
 
<ul>
 
<ul>
−
<li>Choice of the strain of yeast: Sc. M Cherry, wich is resistant to geneticin at a concentration of 200µL/mL</li>
+
<li>Choice of the strain of yeast: Sc. M Cherry, wich is resistant to geneticin at a concentration of 200µL/mL, and produce RFP</li>
 
</ul> <br>
 
</ul> <br>
 
We will have to calculate a survival rate so we have to be able to know how many cells we have in the first place for each experiment:
 
We will have to calculate a survival rate so we have to be able to know how many cells we have in the first place for each experiment:
Line 25: Line 25:
 
<ul>
 
<ul>
 
<li>Centrifuge the culture tube</li>
 
<li>Centrifuge the culture tube</li>
−
<li>Throw the media away, replace it with a known quantity of osmosed water, vortex, take the OD</li>
+
<li>Throw the media away</li>
−
<li>Centrifuge again, throw the water away keeping a very small amount of it to detach the yeasts from the tube easily</li>
+
<li>Add a very small amount 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</li>
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</ul>
 
</ul>
 
<i>Results:</i><br>
 
<i>Results:</i><br>
−
The aluminium is not a good idea, the yeasts stick to it and it gets destroyed when we scratch. We have to find another drying paper.<br>
+
The aluminium is not a good idea, the yeasts stick to it and it gets destroyed when we scratch.<br>
−
 
+
We have to find another drying paper.<br>
 +
<br>
 
<b>Wednesday, Jul. 29</b><br>
 
<b>Wednesday, Jul. 29</b><br>
−
Result of the first OD test:<br>
+
<i>Result of the first OD test:</i><br>
−
The 10mL of solution had an OD of 2,557 and 100µL of the 10^-4 dilution grew 359 colonies.<br>
+
The 10mL solution of yeasts/water had an OD of 2,557 and 100µL of the 10^-4 dilution grew 359 colonies.<br>
 
so '''1 OD = 1,4x10^7 cells/mL'''<br>
 
so '''1 OD = 1,4x10^7 cells/mL'''<br>
 
We launched a similar experience for more repetitions.<br>
 
We launched a similar experience for more repetitions.<br>
 +
<br>
 +
<b>Thursday, Jul. 30</b><br>
 +
<i>Results of the drying:</i><br>
 +
Yeasts grew in the plate. We can check that they produce RFP, it's Sc. M Cherry who survived.<br>
 +
Let's repeat the experience but drying the yeasts on a plastic dish.<br>
 +
This time, we took the OD and plated a known quantity of the obtained powder to be able to calculate the survival rate.<br>
 +
<br>
 +
<b>Friday, Jul. 31</b><br>
 +
<i>Result of the second OD test:</i><br>
 +
<br>
 +
<b>Monday, Aug. 3</b><br>
  
 
<h1>eiuaeuaie</h1>
 
<h1>eiuaeuaie</h1>

Revision as of 12:15, 13 August 2015

Monday Jul. 27
Beginning of the manufacturing project! Let's do it!

  • Choice of the strain of yeast: Sc. M Cherry, wich is resistant to geneticin at a concentration of 200µL/mL, and produce RFP

We will have to calculate a survival rate so we have to be able to know how many cells we have in the first place for each experiment:
  • Centrifuge the culture tube
  • Throw the media away
  • Replace it by a known quantity of osmosed water
  • Take the OD, doing the blank with osmosed water
  • Make dilutions of the yeasts/water solution to plate them
  • Deduce the concentration of Sc. M Cherry corresponding to 1 OD
First try today and plating of the different dilutions on YPD agar+ GEN (200)

Tuesday, Jul. 28
First idea: drying the yeasts.
Protocol:
  • Centrifuge the culture tube
  • Throw the media away
  • Add a very small amount of water to detach the yeasts from the tube easily
  • Spread on aluminium
  • Let it dry for 2 hours
  • Scratch the aluminium to detach the dried yeasts
  • Plate the powder obtained to see if Sc. M Cherry survived
Results:
The aluminium is not a good idea, the yeasts stick to it and it gets destroyed when we scratch.
We have to find another drying paper.

Wednesday, Jul. 29
Result of the first OD test:
The 10mL solution of yeasts/water had an OD of 2,557 and 100µL of the 10^-4 dilution grew 359 colonies.
so '''1 OD = 1,4x10^7 cells/mL'''
We launched a similar experience for more repetitions.

Thursday, Jul. 30
Results of the drying:
Yeasts grew in the plate. We can check that they produce RFP, it's Sc. M Cherry who survived.
Let's repeat the experience but drying the yeasts on a plastic dish.
This time, we took the OD and plated a known quantity of the obtained powder to be able to calculate the survival rate.

Friday, Jul. 31
Result of the second OD test:

Monday, Aug. 3

eiuaeuaie