Team:Cambridge-JIC/Guide/Care/Repropagating
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(Created page with "After transformation, the transformation plates usually contain many transformants, and so growth slows. The plants will usually not form gemmae until they are repropagated. For...") |
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+ | =Repropagating Transformants= | ||
After transformation, the transformation plates usually contain many transformants, and so growth slows. The plants will usually not form gemmae until they are repropagated. | After transformation, the transformation plates usually contain many transformants, and so growth slows. The plants will usually not form gemmae until they are repropagated. | ||
- | For repropagation, use 1/2 B5 + 1.2% agar [+selection if required] plates. Use 5cm deep petri dishes. | + | For repropagation, use [[Team:Cambridge-JIC/Guide/Materials/Media| [1/2 B5 + 1.2% agar [+selection if required] plates]]. Use 5cm, deep petri dishes. |
+ | ==Protocol== | ||
1. Sterilise forceps using ethanol and flame. | 1. Sterilise forceps using ethanol and flame. | ||
2. Working in a laminar flow hood, gently remove a plant (with its rizoids) from the transformation plate and place onto the prepared plates | 2. Working in a laminar flow hood, gently remove a plant (with its rizoids) from the transformation plate and place onto the prepared plates | ||
3. Seal with micropore tape. | 3. Seal with micropore tape. | ||
- | It is recommended to always use freshly poured plates for this stage, to minimize the possibility of contamination. | + | It is recommended to always use freshly autoclaved media and freshly poured plates for this stage, to minimize the possibility of contamination. |
Revision as of 00:52, 17 October 2014
Repropagating Transformants
After transformation, the transformation plates usually contain many transformants, and so growth slows. The plants will usually not form gemmae until they are repropagated.
For repropagation, use [1/2 B5 + 1.2% agar [+selection if required] plates. Use 5cm, deep petri dishes.
Protocol
1. Sterilise forceps using ethanol and flame. 2. Working in a laminar flow hood, gently remove a plant (with its rizoids) from the transformation plate and place onto the prepared plates 3. Seal with micropore tape.
It is recommended to always use freshly autoclaved media and freshly poured plates for this stage, to minimize the possibility of contamination.