Team:Calgary/Notebook/Protocols/catalase

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<p> The purpose of this protocol is to test the ability of KatG-LAA to detoxify hydrogen peroxide. This was important, as catalase will be encorporated into the final sulfur circuit as an optimization feature.
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<li><a href="https://2012.igem.org/Team:Calgary/Notebook/Protocols/onculture">Grow cultures</a> of <i>E. coli J04500-katG+LAA</i> and <i>J04500</i> overnight at 37*C in LB with appropriate antibiotics.</li>
<li><a href="https://2012.igem.org/Team:Calgary/Notebook/Protocols/onculture">Grow cultures</a> of <i>E. coli J04500-katG+LAA</i> and <i>J04500</i> overnight at 37*C in LB with appropriate antibiotics.</li>

Latest revision as of 21:55, 26 October 2012

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Catalase Assay

The purpose of this protocol is to test the ability of KatG-LAA to detoxify hydrogen peroxide. This was important, as catalase will be encorporated into the final sulfur circuit as an optimization feature.

  1. Grow cultures of E. coli J04500-katG+LAA and J04500 overnight at 37*C in LB with appropriate antibiotics.
  2. The following evening, innoculate 20 uL into 3 mL LB with varying concentrations of hydrogen peroxide (0 mM, 1 mM, 5 mM, 10 mM) and grow overnight.
  3. The following morning, observe and record culture turbidity.