Difference between revisions of "Team:Ionis Paris/Protocol 11"
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| + | <a href="#ancre"> <i class="zmdi zmdi-chevron-up btn waves-effect"> </i> </a> | ||
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| − | <h4> | + | <h4>IONIS IGEM</h4> |
| − | <p> | + | <p> We're a group of six different schools from the IONIS Education Group. For this competition we wanted to take advantages of the multiple schools and activity field given by the IONIS education group to create a solid project.</p> |
| − | + | <a href="https://2016.igem.org/Team:Ionis_Paris/Team">Read More</a> | |
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| − | <span>Location: | + | <span>Location: 66 Rue Guy Môquet, 94800 Villejuif, France</span> |
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| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris">Home</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/At_the_lab.html">At the Lab</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/CelloCad"> Side Projects</a></li> |
| − | <li><a href=" | + | <li><a href="Results">Results</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Human_Practice">Human Practice</a></li> |
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Revision as of 12:34, 16 October 2016
Following steps will be performed under a class II PSM Prepare a 50% glycerol stock solution by mixing 5ml glycerol with 5ml distilled, sterile water. Mix 50 µL of the overnight culture of bacteria of interest with 50 µL 50% glycerol solution (25% glycerol final concentration) into a 2 mL cryotube Open the tube and use a sterile loop, toothpick or pipette tip to scrape some of the frozen bacteria off of the top. Do not let the glycerol stock unthaw.
Protocol 11 : Glycerol storage
Aim: Safely store cells at -80°C
Glycerol Freezing Solution (50%)
Mix well to make sure that you see one uniform solution, and there are no layers present.
Bacteria storage
NB : Snap top tubes are not recommended as they can open unexpectedly at -80°C
Label both the lid and the tube of the cryotube. Frozen tubes are hard to write on and samples stored for long periods at -80°C can lose labels stuck to tube.
Store at -80°C
Bacteria recovery
Either streak the bacteria onto an LB agar plate (see culture medium preparation protocol) and grow your bacteria overnight at the appropriate culture condition. The next day you will be able to start an overnight culture for plasmid DNA prep the following day.
Or scrub a bit of bacteria cell and put it into a 500mL sterile Erlenmeyer containing 100mL of sterile liquid LB medium. Cover with aluminum paper and place the Erlenmeyer in a rotative incubator at 37 Celsius degrees, 120rpm.
