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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], 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. | 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 | + | 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. |
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| + | [[File:CAM14_NegativeControl.jpg|thumb|center|300px|Negative control approximately 1.5 weeks after plating]] | ||
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| + | [[Team:Cambridge-JIC/Guide/Constructs/Reporters| Fluorescent]] transformants start appearing from 3-5 days onwards | ||
==Notes== | ==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. | + | - Plates often dry out within 2-3 weeks. You will need to [[Team:Cambridge-JIC/Guide/Care/Repropagating| 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 [[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.
