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Team:Cambridge-JIC/Guide/Transformation/Outline
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| - | Agrobacterium mediated transformation is extensively used in many plant and fungal chassis. For Marchantia, two protocols have been developed to optimise speed and efficiency of transformation | + | Agrobacterium mediated transformation is extensively used in many plant and fungal chassis. For Marchantia, two main protocols have been developed to optimise speed and efficiency of transformation: |
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| + | -the [[Team:Cambridge-JIC/Guide/Transformation/Cocultivation| cocultivation]] method. This requires a shaker capable of shaking a 100ml conical flask at 130rpm under full illumination. | ||
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| + | -the [[Team:Cambridge-JIC/Guide/Transformation/AgarTrap| agar trap]] method. This requires only a ventilated space under full illumination at room temperature. | ||
==Timing== | ==Timing== | ||
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'''For each method, your timing will be optimal if you [[Team:Cambridge-JIC/Guide/Care/PreparingSpores| prepare spores]] and [[Team:Cambridge-JIC/Guide/Transformation/TransformingAgro| transform agrobacteria]] on day 1. | '''For each method, your timing will be optimal if you [[Team:Cambridge-JIC/Guide/Care/PreparingSpores| prepare spores]] and [[Team:Cambridge-JIC/Guide/Transformation/TransformingAgro| transform agrobacteria]] on day 1. | ||
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Latest revision as of 15:34, 17 October 2014
Outline
Agrobacterium mediated transformation is extensively used in many plant and fungal chassis. For Marchantia, two main protocols have been developed to optimise speed and efficiency of transformation:
-the cocultivation method. This requires a shaker capable of shaking a 100ml conical flask at 130rpm under full illumination.
-the agar trap method. This requires only a ventilated space under full illumination at room temperature.
Timing
The cocultivation method requires spores to be (at least) 5 days old prior to cocultivation with agrobacterium, and the agar trap 3 days old. Agrobacteria takes 3 days to grow after transformation.
For each method, your timing will be optimal if you prepare spores and transform agrobacteria on day 1.
