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Team:Cambridge-JIC/Guide/Care/Repropagating
From 2014.igem.org
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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 | + | For repropagation, use [[Team:Cambridge-JIC/Guide/Materials/Media| 1/2 B5 + 1.2% agar plates]], with selection if required. 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. | ||
| + | |||
| + | ''It's OK if you transfer a bit of the agar from the old plate.'' | ||
| + | |||
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. |
| + | ''' | ||
| + | |||
| + | ==Notes== | ||
| + | You can use cuttings of the plant to repropagate it. They will usually grow. Use portions from the edge of the thallus if possible. | ||
| + | <html><br><br><br><br><br><br></html> | ||
Latest revision as of 15:39, 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 plates, with selection if required. 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.
It's OK if you transfer a bit of the agar from the old plate.
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.
Notes
You can use cuttings of the plant to repropagate it. They will usually grow. Use portions from the edge of the thallus if possible.
