Team:Cambridge-JIC/Guide/Transformation/Outline

From 2014.igem.org

Cambridge iGEM 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.