Difference between revisions of "Team:Paris Bettencourt/Notebook/Idli and micro-organisms"
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<h2 class="date one">July 2nd to 9th</h2><br><br> | <h2 class="date one">July 2nd to 9th</h2><br><br> | ||
− | I | + | I experienced different recipes to make Indian idli. I tried to do it with different rice and lentils (called dall in india). Finally, I chose to do |
<a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Idli_recipe">this recipe</a> | <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Idli_recipe">this recipe</a> | ||
with basmati rice and indian dall. <br><br> | with basmati rice and indian dall. <br><br> | ||
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<IMG SRC= "https://static.igem.org/mediawiki/2015/5/52/Idli_water.JPG" width=20%> | <IMG SRC= "https://static.igem.org/mediawiki/2015/5/52/Idli_water.JPG" width=20%> | ||
<br> | <br> | ||
− | I did a media from an idli preparation | + | I did a media from an idli preparation, using 3 glasses of rice (75ml) and 1 glass of dall (25ml) before the soaking phase. After 18 hours of fermentation, I took the water and butter of this idli, mixed it together in a 500ml bottle, and autoclaved it. This media was named "idli water", and it looked like previously. I will just use the clear phase.<br><br> |
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<h2 class="date one">July 12th to 14th and 16th to 18th</h2><br><br> | <h2 class="date one">July 12th to 14th and 16th to 18th</h2><br><br> | ||
− | I | + | I grew ''Saccharomyce cerevisiae'' with mCherry and geneticin resistance genes in idli. I added 2 ml of a yeast growth solution (YPD) with an OD600 of 0.455 (around 1.2 x 10<sup>7</sup> cfu) after the soaking phase and after mixing and grinding the rice (3 glasses of rice ~ 75ml) and dall (1 glass of dall ~ 25ml) together. The first try was to see if there is the fermentation process with these laboratory strains. After the fermentation time, I have plated the µorganisms coming from the butter phase (middle phase between water on the top, and grind matter in the bottom of the erlenmeyer) on YPD with geneticin. I observed a lot of particules of crush rice or dall, and just few colonies, not red like expected with the mCherry gene.<br> |
+ | The second try was more successful. This time I diluted the butter 10 and 100 time before plating, still on YPD and geneticin. I could observed only few colonies for the plate with the 100th and around 30 with the 10th. <br><br> | ||
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<h2 class="date one">July 20th</h2><br><br> | <h2 class="date one">July 20th</h2><br><br> | ||
− | I did a culture on TECAN plate of few species | + | I did a culture on a TECAN plate of few species: ''Saccharomyces cerevisiae'' with mCherry on chromosom, Lactobacillus plantarum NC8 and Lactococcus Lactis MG1363 on their respective media (YPD, MRS and M17), then on "idli water" and on water, with and without antibiotics (respectivly geneticin disulfate, and erythromycin for the two bacteria). <br> |
− | + | We did a TECAN measure after an overnight growth of 18 hours, and we obtained the table followed. We can see that the results are nonsens.<br><br> | |
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<h2 class="date one">August 10th</h2><br><br> | <h2 class="date one">August 10th</h2><br><br> | ||
− | + | We did electrocompetent cells of ''Lactobacillus plantarum'' with <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Electrocompetent_Cells_Preparation_for_Lactobacillus_Plantarum"> a particular protocol</a> <br> | |
It worked very well.<br><br> | It worked very well.<br><br> | ||
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− | + | We did the <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols/Idli_receipe">idli protocol</a>, with 75 ml of rice, for 25 ml of dall. After grinding and mixing, we added 2ml of ''Saccharomyce cerevisiae'' (with mCherry gene and geneticin resistance) that contained 10<sup>8</sup> cells. We plated at t0 and 2h, 4h, 6h, 19h30, 21h30 after t0 three different positons (middle top, lateral middle and bottom middle) at three different concentrations (10<sup>0</sup>, 10<sup>-2</sup> and 10<sup>-3</sup>). <br> | |
<IMG SRC= "https://static.igem.org/mediawiki/2015/2/26/Cfu_number_evolution.png" width=50%> <br> | <IMG SRC= "https://static.igem.org/mediawiki/2015/2/26/Cfu_number_evolution.png" width=50%> <br> | ||
− | + | We can see that the area from where we sample for plating doesn't matter, and the yeast didn't grow very well : just a doubling size of population in 22 hours of culture. <br><br> | |
<div class="column-left"align="center"> | <div class="column-left"align="center"> |
Revision as of 15:35, 14 September 2015