Team:Cambridge-JIC/Guide/Transformation/SelectingTransformants

From 2014.igem.org

Cambridge iGEM 2014


Selecting Transformants

If using the [http://parts.igem.org/Part:BBa_K1484316 Marchantia selection cassette], plate spores onto 1/2 B5 + 100ug/ml cefotaxime + 10ug/ml hygromycin + 1.2% agar plates, and leave under full illumination at room temperature to grow.

Always do a negative control for the transformation to grow. Within 3-5 days, the difference between the transformation plate and the control plate is apparent.

Negative control approximately 1.5 weeks after plating

Fluorescent transformants start appearing from 3-5 days onwards

Notes

- Plates often dry out within 2-3 weeks. You will need to repropagate the transformed plantlets to get continued growth of isogenic gemmae.

- Products of Agrobacterium Mediated Transformation are usually chimeric, since insertions of the T-DNA occur randomly. To get isogenic transformants, you need to harvest gemmae after repropagation.