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Team:Cambridge/Protocols/GetSpores
From 2012.igem.org
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*Pick a single colony from a new plate of B. subtilis and use to inoculate 50ml [[Team:Cambridge/Protocols/SporeMedia|<u><span style="color:#00000CD">CDSM</span></u>]]. | *Pick a single colony from a new plate of B. subtilis and use to inoculate 50ml [[Team:Cambridge/Protocols/SporeMedia|<u><span style="color:#00000CD">CDSM</span></u>]]. | ||
| - | *Grow this for 11 hours in a | + | *Grow this for 11 hours in a 37°C incubator with vigorous aeration. |
| - | *Transfer 2 | + | *Transfer 2.5mL of this to 70ml of new [[Team:Cambridge/Protocols/SporeMedia|<u><span style="color:#00000CD">CDSM</span></u>]] culture. Incubate new CDSM culture and original 50mL CDSM culture for a further 37hrs, to a total of 48hrs. |
*Longer incubation may be needed for full release of spores. | *Longer incubation may be needed for full release of spores. | ||
{{Template:Team:Cambridge/CAM_2012_TEMPLATE_FOOT}} | {{Template:Team:Cambridge/CAM_2012_TEMPLATE_FOOT}} | ||
Revision as of 16:51, 19 September 2012
Producing high yields of B.subtilis spores
Preparation
Plate and grow B. subtilis overnight on antibiotic resistant plates.
Method
- Pick a single colony from a new plate of B. subtilis and use to inoculate 50ml CDSM.
- Grow this for 11 hours in a 37°C incubator with vigorous aeration.
- Transfer 2.5mL of this to 70ml of new CDSM culture. Incubate new CDSM culture and original 50mL CDSM culture for a further 37hrs, to a total of 48hrs.
- Longer incubation may be needed for full release of spores.
