Difference between revisions of "Team:Ionis Paris/Protocol 6"
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| − | <h4>IONIS | + | <h4>iGEM IONIS</h4> |
| − | <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> | + | <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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<a href="mailto:igem.ionis@gmail.com">email: igem.ionis@gmail.com</a> | <a href="mailto:igem.ionis@gmail.com">email: igem.ionis@gmail.com</a> | ||
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| − | <li><a href="https://2016.igem.org/ | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Working_at_La_Paillasse">in the Lab</a></li> |
| − | <li><a href="https://2016.igem.org/ | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Measurement"> Side Projects</a></li> |
| − | <li><a href=" | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Parts">Results</a></li> |
| − | <li><a href="https://2016.igem.org/Team:Ionis_Paris/ | + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/HPintro">Human |
| − | <li><a href="https://2016.igem.org/Team | + | Practice</a></li> |
| + | <li><a href="https://2016.igem.org/Team:Ionis_Paris/Team">Team</a></li> | ||
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| − | <p>©IONIS_IGEM_2016</a>.</p> | + | <p>©IONIS_IGEM_2016</a>.</p> |
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Revision as of 11:02, 18 October 2016
Thaw a tube of DH5-alpha competent cells on ice for 10 min.
Protocol 6 : E.Coli Dh5α and BL21 transformation (Heat-shock)
Aim: Introduce plasmid DNA in E.Coli bacteria
Mix gently and carefully pipette 50 µL of cells into a transformation tube on ice
Add 1-5µL plasmid DNA to the cell mixture (one tube for each ratio)
Carefully flick the tube 4-5 times to mix cells and DNA. Do not vortex.
Place the mixture on ice for 30 min. Do not mix.
Heat shock at exactly 42°C for 45 s. Do not mix.
Place on ice for 5 min. Do not mix.
Pipette 250µL of room temperature SOC into the mixture.
Place at 37°C for 60 min at 250 rpm.
Warm selection plates to 25°C.
Mix the cells by flicking the tube and inverting.
Spread 100µL of cells onto selection plates.
Incubate all the plates O/N at 37°C.
