Team:Caltech/Protocols/H2O2 Production Assay

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# 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.
# 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.
# In the morning, dilute culture 1:100 into 5 ml SOC +Amp +IPTG in triplicate.
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# Grow culures to an OD600 of ~0.8, then [[Team:Caltech/Protocols/Measuring_H2O2|assay supernatant]] for hydrogen peroxide approximately every 30 minutes.
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# Grow cultures to an OD600 of ~0.8, then [[Team:Caltech/Protocols/Measuring_H2O2|assay supernatant]] for hydrogen peroxide approximately every 30 minutes.
# 1hr after reaching an OD600 of ~0.8, induce cultures with 10 nM AHL.
# 1hr after reaching an OD600 of ~0.8, induce cultures with 10 nM AHL.
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H2O2 Production Assay


  1. Grow an overnight culture of JI377 transformed with K137076 or luxR (negative control) on pSB1A2 in SOC + Amp.
  2. In the morning, dilute culture 1:100 into 5 ml SOC +Amp +IPTG in triplicate.
  3. Grow cultures to an OD600 of ~0.8, then assay supernatant for hydrogen peroxide approximately every 30 minutes.
  4. 1hr after reaching an OD600 of ~0.8, induce cultures with 10 nM AHL.
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