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Team:Cambridge-JIC/Guide/Transformation/SelectingTransformants
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=Selecting Transformants= | =Selecting Transformants= | ||
| - | If using the [http://parts.igem.org/Part:BBa_K1484316 Marchantia selection cassette | + | If using the [http://parts.igem.org/Part:BBa_K1484316 Marchantia selection cassette], plate spores onto [[Team:Cambridge-JIC/Guide/Materials/Media | 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 appear from 3 days onwards. | 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 appear from 3 days onwards. | ||
| - | Plates often dry out within 2-3 weeks. You will need to [[Team:Cambridge-JIC/Guide/Care/Repropagating| repropagate]] the transformed plantlets to get growth of isogenic gemmae. | + | ==Notes== |
| + | - Plates often dry out within 2-3 weeks. You will need to [[Team:Cambridge-JIC/Guide/Care/Repropagating| repropagate]] the transformed plantlets to get 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 [[Team:Cambridge-JIC/Guide/Care/HarvestingGemmae| harvest gemmae]] after repropagation. | ||
Revision as of 00:56, 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 appear from 3 days onwards.
Notes
- Plates often dry out within 2-3 weeks. You will need to repropagate the transformed plantlets to get 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.
