Difference between revisions of "Team:Paris Bettencourt/Notebook/VitaminB12"
(15 intermediate revisions by the same user not shown) | |||
Line 1: | Line 1: | ||
{{Paris_Bettencourt/header}} | {{Paris_Bettencourt/header}} | ||
{{Paris_Bettencourt/menu}} | {{Paris_Bettencourt/menu}} | ||
− | {{Paris_Bettencourt/ | + | {{Paris_Bettencourt/banner|page_id=notebook|page_name=Notebook - Vitamin B12}} |
<html> | <html> | ||
Line 7: | Line 7: | ||
<div id="notebookMenu"> | <div id="notebookMenu"> | ||
<ul> | <ul> | ||
− | <li><a href="#july">July</a></li> | + | <li class="stretch"><a href="#july">July</a></li> |
<li><a href="#august">August</a></li> | <li><a href="#august">August</a></li> | ||
<li><a href="#september">September</a></li> | <li><a href="#september">September</a></li> | ||
Line 27: | Line 27: | ||
<tr><td>Yeast extract</td><td>5 g</td></tr> | <tr><td>Yeast extract</td><td>5 g</td></tr> | ||
<tr><td>Glucose</td><td>3 g</td></tr> | <tr><td>Glucose</td><td>3 g</td></tr> | ||
− | <tr><td>Water</td><td>Fill to | + | <tr><td>Water</td><td>Fill to 1 L</td></tr> |
</table> | </table> | ||
</li> | </li> | ||
Line 50: | Line 50: | ||
<h2 class="date one">July 30th</h2> | <h2 class="date one">July 30th</h2> | ||
− | <img style="float:right" src="https://static.igem.org/mediawiki/2015/f/f7/PB_B12_Result_plates.png" width="400px"> | + | <div style="float: right; width: 400px;"><img style="float:right" src="https://static.igem.org/mediawiki/2015/f/f7/PB_B12_Result_plates.png" width="400px"><br> |
+ | <b>Figure 1:</b> Result of the growth of different strains on different media.</div> | ||
<ul> | <ul> | ||
<li>Tested the YEL+agar medium with the following strains: | <li>Tested the YEL+agar medium with the following strains: | ||
Line 60: | Line 61: | ||
<li><i>Saccharomyces cerevisiae</i></li> | <li><i>Saccharomyces cerevisiae</i></li> | ||
</ul></li> | </ul></li> | ||
− | <li>Result (after 24 hours) | + | <li>Result (after 24 hours): figure 1.</li> |
</ul> | </ul> | ||
Line 84: | Line 85: | ||
<h2 class="date two">August 15th</h2> | <h2 class="date two">August 15th</h2> | ||
<ul> | <ul> | ||
− | <li>Successfully grew <i>P. freudenreichii</i> in | + | <li>Successfully grew <i>P. freudenreichii</i> in MEA liquid and YEL liquid</li> |
<li>It seemed to grow much faster. Do I have a contanimation? Need to check.</li> | <li>It seemed to grow much faster. Do I have a contanimation? Need to check.</li> | ||
</ul> | </ul> | ||
Line 136: | Line 137: | ||
</ul> | </ul> | ||
− | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <a name="september" class="anchor"><h1></h1></a> | ||
+ | |||
+ | |||
+ | <h2 class="date three">September 1st</h2> | ||
+ | <div style="width: 300px; float: right"><img src="https://static.igem.org/mediawiki/2015/b/b6/PB_Ewen_16PCRGel01.png" width="160px"><br> | ||
+ | <b>Figure 2: </b> Gel result of the 16S rRNA PCR.<br><b>Lane 10:</b> PCR on the 10 ng/μl of extracted DNA, <b>Lane 5:</b> PCR on the 5 ng/μl of extracted DNA, <b>+:</b> positive control that should have worked.</div> | ||
+ | <ul> | ||
+ | <li>Made a 1 M Tris solution, adjusted the pH to 8.3</li> | ||
+ | <li>Remade an enzymatic lysis buffer, with the right reagents this time.</li> | ||
+ | <li>Tried again the <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#QiagenDNeasy">DNeasy extraction kit</a>. Got the following yield: | ||
+ | <table> | ||
+ | <tr><th>Start volume</th><th>DNA extracted</th></tr> | ||
+ | <tr><td>1 ml</td><td>10.3 ng/μl</td></tr> | ||
+ | <tr><td>0.5 ml</td><td>5.2 ng/μl</td></tr> | ||
+ | </table><br> | ||
+ | The yield is better, will try a PCR on it. | ||
+ | </li> | ||
+ | <li>Made a PCR following this <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#FSPCR">protocol</a> and using the following primers: | ||
+ | <table> | ||
+ | <tr><th>Name</th><th>Sequence</th></tr> | ||
+ | <tr><td>16s universal primer <b>27F</b></td><td>AGA GTT TGA TCM TGG CTC AG</td></tr> | ||
+ | <tr><td>16s universal primer <b>1492R</b></td><td>CGG TTA CCT TGT TAC GAC TT</td></tr> | ||
+ | </table> | ||
+ | </li> | ||
+ | <li>Result on Figure 2.</li> | ||
+ | <li>The Mastermix was probably not working.</li> | ||
+ | </ul> | ||
+ | <div style="clear:both"></div> | ||
+ | |||
+ | |||
+ | <h2 class="date three">September 8th</h2> | ||
+ | <ul> | ||
+ | <div style="float:right; width: 300px;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2015/1/1a/PB_Ewen_gelJason.png" width="100px"><br> | ||
+ | <b>Figure 3:</b> Gel result of the 16S PCR. <b>"-" lane:</b>Negative control, <b>"x" lane:</b>PCR on the 10 ng/μl. | ||
+ | </div> | ||
+ | <li>Tried again a <a href="https://2015.igem.org/Team:Paris_Bettencourt/Protocols#FSPCR">PCR</a> with the same primers, but a new mastermix.</li> | ||
+ | <li>Result: figure 3. It worked!</li> | ||
+ | </ul> | ||
+ | <div style="clear:both"></div> | ||
Latest revision as of 22:14, 18 September 2015