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| - | An overnight culture of JI377 transformed with luxR+GFP or luxR (negative control) on pSB1A2 in SOC +Amp was back diluted 1:100 into 5 ml SOC +Amp +IPTG in triplicate. The cultures were grown to an OD600 of ~0.8, at which point the supernatant was assayed for hydrogen peroxide approximately every 30 minutes. 1hr after reaching an OD600 of ~0.8, the cultures were induced with 10 nM AHL.
| + | # Grow an overnight culture of JI377 transformed with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K137076 K137076] or [http://partsregistry.org/wiki/index.php?title=Part:BBa_F2622 luxR] (negative control) on pSB1A2 in SOC + Amp. |
| | + | # In the morning, dilute culture 1:100 into 5 ml SOC +Amp +IPTG in triplicate. |
| | + | # Grow culures to an OD600 of ~0.8, then assay supernatant for hydrogen peroxide approximately every 30 minutes. |
| | + | # 1hr after reaching an OD600 of ~0.8, induce cultures with 10 nM AHL. |
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Revision as of 22:05, 17 October 2008
iGEM 2008
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H2O2 Production Assay
- Grow an overnight culture of JI377 transformed with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K137076 K137076] or [http://partsregistry.org/wiki/index.php?title=Part:BBa_F2622 luxR] (negative control) on pSB1A2 in SOC + Amp.
- In the morning, dilute culture 1:100 into 5 ml SOC +Amp +IPTG in triplicate.
- Grow culures to an OD600 of ~0.8, then assay supernatant for hydrogen peroxide approximately every 30 minutes.
- 1hr after reaching an OD600 of ~0.8, induce cultures with 10 nM AHL.
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