Difference between revisions of "Team:Freiburg/Notebook"

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{{Freiburg}}
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{{Freiburg/Main}}
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{{Freiburg/Menubar}}
 
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<html>
  
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<h1 class="sectionedit1"><a name="sporulation" id="sporulation">Sporulation</a></h1>
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<div class="level1">
  
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<div class="column full_size">
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<p>
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<strong>Checklist:</strong><br/>
  
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<p> Document the dates you worked on your project.</p>
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</p>
 +
<ul>
 +
<li class="level1"><div class="li"> Use the right Medium.</div>
 +
</li>
 +
<li class="level1"><div class="li"> Vortex cuvettes if you make a dilution</div>
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</li>
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<li class="level1"><div class="li"> Set Nanodrop to cuvettes</div>
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</li>
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</ul>
  
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</div>
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<p>
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1. Overnight culture
 +
</p>
 +
<ul>
 +
<li class="level1"><div class="li"> inoculate your culture in 5ml LB-Medium</div>
 +
</li>
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<li class="level1"><div class="li"> let them grow over night at 37°C, 200rpm</div>
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</li>
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</ul>
  
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<div class="column half_size">
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<p>
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<h5>What should this page have?</h5>
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<br/>
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2. Exponential growth
 +
</p>
 
<ul>
 
<ul>
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<li>Chronological notes of what your team is doing.</li>
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<li class="level1"><div class="li"> messure the OD600 of your overnight culture</div>
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<li> Brief descriptions of daily important events.</li>
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</li>
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<li>Pictures of your progress. </li>
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<li class="level1"><div class="li"> inoculate 2ml + Xml to measure OD (personal preference) LB-Medium to OD600=0,1 from overnight culture </div>
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<li>Mention who participated in what task.</li>
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</li>
 +
<li class="level1"><div class="li"> let the cells grow to an OD600=0,8 (37°C, 200rpm)</div>
 +
</li>
 
</ul>
 
</ul>
 +
 +
<p>
 +
3. Sporulation
 +
</p>
 +
<ul>
 +
<li class="level1"><div class="li"> centrifuge the 2 ml of cells at 13.000 rpm for 1 minute</div>
 +
</li>
 +
<li class="level1"><div class="li"> wash the pellet with PBS</div>
 +
</li>
 +
<li class="level1"><div class="li"> resuspend the pellet in 1 ml DSM (Difco Sporulation Medium)<a href="/igem2016/doku.php?id=labprotocols:media" class="wikilink1" title="labprotocols:media">Culture medium</a></div>
 +
</li>
 +
<li class="level1"><div class="li"> let the cells grow (24h,37°C 200rpm)</div>
 +
</li>
 +
</ul>
 +
 +
<p>
 +
4. Lysozym treatment
 +
</p>
 +
<ul>
 +
<li class="level1"><div class="li"> dilute the cells 6:1 in lysozym solution (15mg/μl)</div>
 +
</li>
 +
<li class="level1"><div class="li"> incubate for 1 hour at room temperature</div>
 +
</li>
 +
<li class="level1"><div class="li"> wash 6 times with 1x PBS</div>
 +
</li>
 +
</ul>
 +
 +
<p>
 +
5. Count the spores (Neubauer-counting chamber, usually dilute in BPS 1 to 100)<br/>
 +
 +
<a href="/igem2016/lib/exe/fetch.php?media=labprotocols:sorecountingb54.png" class="media" title="labprotocols:sorecountingb54.png"><img src="/igem2016/lib/exe/fetch.php?w=400&amp;media=labprotocols:sorecountingb54.png" class="media" alt="" width="400" /></a><br/>
 +
 +
6. Make aliquots (around 100million spores per aliquot)
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</p>
  
 
</div>
 
</div>
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<!-- EDIT1 SECTION "Sporulation" [1-1087] -->
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<h1 class="sectionedit2"><a name="induktion_of_sporulation" id="induktion_of_sporulation">Induktion of sporulation</a></h1>
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<div class="level1">
  
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<div class="column half_size">
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<p>
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<h5>Inspiration</h5>
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<a href="https://www.researchgate.net/post/How_to_induce_sporulation_of_Bacillus_spp_by_heating_or_any_other_simple_technique" class="urlextern" target="_Blank" title="https://www.researchgate.net/post/How_to_induce_sporulation_of_Bacillus_spp_by_heating_or_any_other_simple_technique"  rel="nofollow">https://www.researchgate.net/post/How_to_induce_sporulation_of_Bacillus_spp_by_heating_or_any_other_simple_technique</a><br/>
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<p>You can see what others teams have done to organize their notes:</p>
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<ul>
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<a href="https://www.researchgate.net/post/How_to_sporulate_the_bacteria_or_how_to_induce_the_sporulation_in_bacteria_especially_for_Bacillus_spp" class="urlextern" target="_Blank" title="https://www.researchgate.net/post/How_to_sporulate_the_bacteria_or_how_to_induce_the_sporulation_in_bacteria_especially_for_Bacillus_spp" rel="nofollow">https://www.researchgate.net/post/How_to_sporulate_the_bacteria_or_how_to_induce_the_sporulation_in_bacteria_especially_for_Bacillus_spp</a><br/>
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<li><a href="https://2014.igem.org/Team:ATOMS-Turkiye/Notebook">2014 ATOMS-Turkiye</a></li>
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<li><a href="https://2014.igem.org/Team:Tec-Monterrey/ITESM14_project.html#tab_notebook">2014 Tec Monterrey</a></li>
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</p>
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<li><a href="https://2014.igem.org/Team:Kyoto/Notebook/Magnetosome_Formation#title">2014 Kyoto</a></li>
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<div class="tags"><span>
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<li><a href="https://2014.igem.org/Team:Cornell/notebook">2014 Cornell</a></li>
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<a href="/igem2016/doku.php?id=tag:workwithbs_labprotocols" class="wikilink1" title="tag:workwithbs_labprotocols" rel="tag">WorkwithBS labprotocols</a>
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</ul>
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</span></div>
  
 
</div>
 
</div>
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<!-- EDIT2 SECTION "Induktion of sporulation" [1088-] -->
 
</html>
 
</html>

Revision as of 11:40, 15 August 2016

Menu

Sporulation

Checklist:

  • Use the right Medium.
  • Vortex cuvettes if you make a dilution
  • Set Nanodrop to cuvettes

1. Overnight culture

  • inoculate your culture in 5ml LB-Medium
  • let them grow over night at 37°C, 200rpm


2. Exponential growth

  • messure the OD600 of your overnight culture
  • inoculate 2ml + Xml to measure OD (personal preference) LB-Medium to OD600=0,1 from overnight culture
  • let the cells grow to an OD600=0,8 (37°C, 200rpm)

3. Sporulation

  • centrifuge the 2 ml of cells at 13.000 rpm for 1 minute
  • wash the pellet with PBS
  • resuspend the pellet in 1 ml DSM (Difco Sporulation Medium)Culture medium
  • let the cells grow (24h,37°C 200rpm)

4. Lysozym treatment

  • dilute the cells 6:1 in lysozym solution (15mg/μl)
  • incubate for 1 hour at room temperature
  • wash 6 times with 1x PBS

5. Count the spores (Neubauer-counting chamber, usually dilute in BPS 1 to 100)

6. Make aliquots (around 100million spores per aliquot)

Induktion of sporulation

https://www.researchgate.net/post/How_to_induce_sporulation_of_Bacillus_spp_by_heating_or_any_other_simple_technique
https://www.researchgate.net/post/How_to_sporulate_the_bacteria_or_how_to_induce_the_sporulation_in_bacteria_especially_for_Bacillus_spp