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Team:Cambridge-JIC/Guide/Transformation/PreparingElectrocompetentAgro
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| - | Many protocols for preparing electrocompetent agrobacteria can be found online, and are roughly the same. The protocol given | + | Many protocols for preparing electrocompetent agrobacteria can be found online, and are roughly the same. The [[Team:Cambridge-JIC/Guide/Transformation/TransformingAgro | |protocol]] given in this guide for electroporation uses 50ul of agrobacteria cells per transformation- so you should aliquot appropriately. |
For example: | For example: | ||
Revision as of 00:17, 17 October 2014
Many protocols for preparing electrocompetent agrobacteria can be found online, and are roughly the same. The |protocol given in this guide for electroporation uses 50ul of agrobacteria cells per transformation- so you should aliquot appropriately.
For example: http://www.plantsci.cam.ac.uk/research/jillharrison/protocols/cloning/preparation-of-agrobacterium-electrocompetent.pdf
Although electrocompetent agrobacteria are available for purchase, you will want to create a stock of electrocompetent agrobacteria containing the Ti plasmid, since double transformation can lead to low efficiencies.
After you've prepared these cells, plate an untransformed batch onto selective plates with the antibiotic you are planning on transforming them with. Make sure that no colonies appear!
See transforming Agrobacterium for the next steps.
