Team:LCG-UNAM-Mexico/Resources/Protocols/Transformation

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(New page: Transformation<br> Take 50micro liters of chemically competent cells (DH5alpha in our case) stored into a fridge at -70°C. Put the cells in ice. <br> Add the plasmid to be transformed (T...)
 
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Transformation<br>
Transformation<br>
Take 50micro liters of chemically competent cells (DH5alpha in our case) stored into a fridge at -70°C.  
Take 50micro liters of chemically competent cells (DH5alpha in our case) stored into a fridge at -70°C.  
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Discard the supernatant.
Discard the supernatant.
Plate into a petri box.
Plate into a petri box.
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Latest revision as of 19:12, 21 October 2009


Transformation
Take 50micro liters of chemically competent cells (DH5alpha in our case) stored into a fridge at -70°C. Put the cells in ice.
Add the plasmid to be transformed (The quantity is depending of the source and concentration).
Let the samples in ice for 20 minutes.
Apply a heat shock at 41°C for 1 minute.
Incubate on ice for 5 minutes.
Incorporate and mix well 1mL of liquid-LB medium.
Mix at 250RPM for 1 hour.
Centrifugue to concentrate for 1 minute at 13000RPM. Discard the supernatant. Plate into a petri box.


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