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Team:Cambridge-JIC/Guide/Transformation/SelectingTransformants
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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. | 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. | ||
| - | [[File: | + | [[File:CAM14_NegativeControl.jpg|thumb|center|300px|Negative control approximately 1.5 weeks after plating]] |
[[Team:Cambridge-JIC/Guide/Constructs/Reporters| Fluorescent]] transformants start appearing from 3-5 days onwards | [[Team:Cambridge-JIC/Guide/Constructs/Reporters| Fluorescent]] transformants start appearing from 3-5 days onwards | ||
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- Products of Agrobacterium Mediated Transformation are usually chimeric, since insertions of the T-DNA occur randomly. To get isogenic transformants, you need to [[Team:Cambridge-JIC/Guide/Care/HarvestingGemmae| harvest gemmae]] after repropagation. | - Products of Agrobacterium Mediated Transformation are usually chimeric, since insertions of the T-DNA occur randomly. To get isogenic transformants, you need to [[Team:Cambridge-JIC/Guide/Care/HarvestingGemmae| harvest gemmae]] after repropagation. | ||
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Latest revision as of 15:37, 17 October 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.
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.
