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Team:Cambridge-JIC/Guide/Materials/Media
From 2014.igem.org
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- Agrobacterium plates | - Agrobacterium plates | ||
| + | Use standard LB + 1.5% agar + required antibiotics. | ||
- Marchantia plates | - Marchantia plates | ||
| + | For marchantia, 1/2 Gamborg's B5 media + 1.2% agar is effective. Gamborg's B5 salts can be found readily from chemical suppliers. | ||
| + | |||
| + | Always use freshly autoclaved media, and work in a laminar flow hood, maintaining sterile conditions. | ||
| + | |||
| + | Fungal spores rapidly colonise the plates under conditions for Marchantia growth. | ||
| + | |||
| + | Always seal all plates using micropore tape. | ||
- Selection plates | - Selection plates | ||
| + | If using the Marchantia polymorpha resistance cassette and agrobacterium mediated transformation, use Marchantia plates with 10ug/ml hygromycin and 100ug/ml cefotaxime (to ensure that agrobacterium does not colonize the plate). | ||
| + | |||
| + | Note: selection plates do not contain a high enough concentration of hygromycin to preclude fungal colonisation. Work in sterile conditions. | ||
| + | |||
- Acetosyringone | - Acetosyringone | ||
| + | Use a fresh stock, with a concentration of 100mM, in DMSO. This will be used in transformation protocols as 1000x. | ||
Revision as of 21:20, 16 October 2014
- Agrobacterium plates Use standard LB + 1.5% agar + required antibiotics. - Marchantia plates For marchantia, 1/2 Gamborg's B5 media + 1.2% agar is effective. Gamborg's B5 salts can be found readily from chemical suppliers.
Always use freshly autoclaved media, and work in a laminar flow hood, maintaining sterile conditions.
Fungal spores rapidly colonise the plates under conditions for Marchantia growth.
Always seal all plates using micropore tape. - Selection plates If using the Marchantia polymorpha resistance cassette and agrobacterium mediated transformation, use Marchantia plates with 10ug/ml hygromycin and 100ug/ml cefotaxime (to ensure that agrobacterium does not colonize the plate).
Note: selection plates do not contain a high enough concentration of hygromycin to preclude fungal colonisation. Work in sterile conditions.
- Acetosyringone Use a fresh stock, with a concentration of 100mM, in DMSO. This will be used in transformation protocols as 1000x.
