Team:Cambridge/Protocols/GetSpores

From 2012.igem.org

(Difference between revisions)
(Method)
(Method)
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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>]].
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*Grow this for 11 hours in a 37<sup>o</sup>C incubator with vigorous aeration.
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*Grow this for 11 hours in a 37°C incubator with vigorous aeration.
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*Transfer 2-5% of this to make up another 50ml volume of [[Team:Cambridge/Protocols/SporeMedia|<u><span style="color:#00000CD">CDSM</span></u>]] culture and grow both for a total incubation time of 48 hours.
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*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.